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Sample records for buds

  1. Axillary bud development in chrysanthemum

    OpenAIRE

    Ruiter, de, H.

    1996-01-01


    Each chrysanthemum cutting originates from an axillary bud. For an improvement of the cultivation of cuttings or more specific their quality, it is necessary that the development of an axillary bud can be controlled as good as possible. Axillary bud development can be distinguished into axillary bud formation and axillary bud outgrowth. The effect of assimilates, position and age of axillary buds, and temperature on formation and outgrowth of the axillary buds and the subsequent cu...

  2. What Are Taste Buds?

    Science.gov (United States)

    ... for Kids? Your Teeth Heart Murmurs What Are Taste Buds? KidsHealth > For Kids > What Are Taste Buds? Print A A A en español ¿Qué ... Did you ever wonder why your favorite foods taste so good? Well, you can thank your taste ...

  3. "Bud, Not Buddy."

    Science.gov (United States)

    Brodie, Carolyn S.

    2002-01-01

    Discusses the award-winning book "Bud, Not Buddy" written by Christopher Paul Curtis. Lists different versions of the book; suggests learning activities; lists sources for biographical information and interviews with Curtis, teacher guides, professional articles, and other Depression era novels; and provides a citation for the author's…

  4. Axillary bud development in chrysanthemum

    NARCIS (Netherlands)

    Ruiter, de H.A.

    1996-01-01


    Each chrysanthemum cutting originates from an axillary bud. For an improvement of the cultivation of cuttings or more specific their quality, it is necessary that the development of an axillary bud can be controlled as good as possible. Axillary bud development can be distinguished into

  5. Axillary bud development in rose

    OpenAIRE

    Marcelis - van Acker, C.A.M.

    1994-01-01

    Axillary buds form the basis of flower production of a rose crop. Within a rose crop there exists an undesired large variation in shoot number and size, which affects flower yield. Part of this variation may be traced back to early variation in axillary buds. The aim of the research reported in this thesis was to enlarge the knowledge and insight in the development of axillary buds. It was investigated to what extent the growth of an axillary bud into a shoot can be influenced during...

  6. Axillary bud development in rose

    NARCIS (Netherlands)

    Marcelis - van Acker, C.A.M.

    1994-01-01

    Axillary buds form the basis of flower production of a rose crop. Within a rose crop there exists an undesired large variation in shoot number and size, which affects flower yield. Part of this variation may be traced back to early variation in axillary buds. The aim of the research

  7. Tumor budding in colorectal carcinomas.

    Science.gov (United States)

    Sert Bektaş, Sevda; Inan Mamak, Gülsün; Cırış, Ibrahim Metin; Bozkurt, Kemal Kürşat; Kapucuoğlu, Nilgün

    2012-01-01

    In colorectal carcinomas, tumor budding has been defined as the presence of isolated single tumor cells or small cell clusters in the stroma at the invasive tumor margin. In this study, the relationship between tumor budding density at the invasive tumor margin and pathological parameters is investigated. Haematoxylin and eosin stained slides of 73 cases with colorectal carcinoma were retrospectively evaluated for the presence and intensity of tumor budding by 2 observers. After the specimens were assessed, the highest density of tumor budding area was counted in a microscopic field of x200. Cases were separated into 2 groups according to tumor budding density as low grade ( tumor invasion, histological grade, vascular invasion and lymph node involvement was investigated. Of the 73 colorectal carcinoma cases, 33 (45.2%) had low and 40 (54.8%) had high grade tumor budding density, respectively. There was a statistically significant relationship between high grade tumor budding density and histological grade (p=0.042), lymph node involvement (p=0.0001) and vascular invasion (p=0.0034). High grade tumor budding density is associated with aggressive phenotypical features in colorectal carcinoma.

  8. Coevolutionary patterning of teeth and taste buds

    OpenAIRE

    Bloomquist, R.F.; Parnell, N; Phillips, K A; Fowler, T E; Yu, Tian; Sharpe, Paul T.; Streelman, J T

    2015-01-01

    Teeth and taste buds are iteratively patterned structures that line the oro-pharynx of vertebrates. Biologists do not fully understand how teeth and taste buds develop from undifferentiated epithelium or how variation in organ density is regulated. These organs are typically studied independently because of their separate anatomical location in mammals: teeth on the jaw margin and taste buds on the tongue. However, in many aquatic animals like bony fishes, teeth and taste buds are colocalized...

  9. Foamy Virus Budding and Release

    Directory of Open Access Journals (Sweden)

    Dirk Lindemann

    2013-04-01

    Full Text Available Like all other viruses, a successful egress of functional particles from infected cells is a prerequisite for foamy virus (FV spread within the host. The budding process of FVs involves steps, which are shared by other retroviruses, such as interaction of the capsid protein with components of cellular vacuolar protein sorting (Vps machinery via late domains identified in some FV capsid proteins. Additionally, there are features of the FV budding strategy quite unique to the spumaretroviruses. This includes secretion of non-infectious subviral particles and a strict dependence on capsid-glycoprotein interaction for release of infectious virions from the cells. Virus-like particle release is not possible since FV capsid proteins lack a membrane-targeting signal. It is noteworthy that in experimental systems, the important capsid-glycoprotein interaction could be bypassed by fusing heterologous membrane-targeting signals to the capsid protein, thus enabling glycoprotein-independent egress. Aside from that, other systems have been developed to enable envelopment of FV capsids by heterologous Env proteins. In this review article, we will summarize the current knowledge on FV budding, the viral components and their domains involved as well as alternative and artificial ways to promote budding of FV particle structures, a feature important for alteration of target tissue tropism of FV-based gene transfer systems.

  10. Development and growth potential of axillary buds in roses as affected by bud age.

    NARCIS (Netherlands)

    Marcelis-van Acker, C.A.M.

    1994-01-01

    The effect of axillary bud age on the development and potential for growth of the bud into a shoot was studied in roses. Age of the buds occupying a similar position on the plant varied from 'subtending leaf just unfolded' up to 1 year later. With increasing age of the axillary bud its dry mass,

  11. Assembly and budding of Ebolavirus.

    Directory of Open Access Journals (Sweden)

    Takeshi Noda

    2006-09-01

    Full Text Available Ebolavirus is responsible for highly lethal hemorrhagic fever. Like all viruses, it must reproduce its various components and assemble them in cells in order to reproduce infectious virions and perpetuate itself. To generate infectious Ebolavirus, a viral genome-protein complex called the nucleocapsid (NC must be produced and transported to the cell surface, incorporated into virions, and then released from cells. To further our understanding of the Ebolavirus life cycle, we expressed the various viral proteins in mammalian cells and examined them ultrastructurally and biochemically. Expression of nucleoprotein alone led to the formation of helical tubes, which likely serve as a core for the NC. The matrix protein VP40 was found to be critical for transport of NCs to the cell surface and for the incorporation of NCs into virions, where interaction between nucleoprotein and the matrix protein VP40 is likely essential for these processes. Examination of virus-infected cells revealed that virions containing NCs mainly emerge horizontally from the cell surface, whereas empty virions mainly bud vertically, suggesting that horizontal budding is the major mode of Ebolavirus budding. These data form a foundation for the identification and development of potential antiviral agents to combat the devastating disease caused by this virus.

  12. Assembly and budding of Ebolavirus.

    Science.gov (United States)

    Noda, Takeshi; Ebihara, Hideki; Muramoto, Yukiko; Fujii, Ken; Takada, Ayato; Sagara, Hiroshi; Kim, Jin Hyun; Kida, Hiroshi; Feldmann, Heinz; Kawaoka, Yoshihiro

    2006-09-01

    Ebolavirus is responsible for highly lethal hemorrhagic fever. Like all viruses, it must reproduce its various components and assemble them in cells in order to reproduce infectious virions and perpetuate itself. To generate infectious Ebolavirus, a viral genome-protein complex called the nucleocapsid (NC) must be produced and transported to the cell surface, incorporated into virions, and then released from cells. To further our understanding of the Ebolavirus life cycle, we expressed the various viral proteins in mammalian cells and examined them ultrastructurally and biochemically. Expression of nucleoprotein alone led to the formation of helical tubes, which likely serve as a core for the NC. The matrix protein VP40 was found to be critical for transport of NCs to the cell surface and for the incorporation of NCs into virions, where interaction between nucleoprotein and the matrix protein VP40 is likely essential for these processes. Examination of virus-infected cells revealed that virions containing NCs mainly emerge horizontally from the cell surface, whereas empty virions mainly bud vertically, suggesting that horizontal budding is the major mode of Ebolavirus budding. These data form a foundation for the identification and development of potential antiviral agents to combat the devastating disease caused by this virus.

  13. Coevolutionary patterning of teeth and taste buds.

    Science.gov (United States)

    Bloomquist, Ryan F; Parnell, Nicholas F; Phillips, Kristine A; Fowler, Teresa E; Yu, Tian Y; Sharpe, Paul T; Streelman, J Todd

    2015-11-03

    Teeth and taste buds are iteratively patterned structures that line the oro-pharynx of vertebrates. Biologists do not fully understand how teeth and taste buds develop from undifferentiated epithelium or how variation in organ density is regulated. These organs are typically studied independently because of their separate anatomical location in mammals: teeth on the jaw margin and taste buds on the tongue. However, in many aquatic animals like bony fishes, teeth and taste buds are colocalized one next to the other. Using genetic mapping in cichlid fishes, we identified shared loci controlling a positive correlation between tooth and taste bud densities. Genome intervals contained candidate genes expressed in tooth and taste bud fields. sfrp5 and bmper, notable for roles in Wingless (Wnt) and bone morphogenetic protein (BMP) signaling, were differentially expressed across cichlid species with divergent tooth and taste bud density, and were expressed in the development of both organs in mice. Synexpression analysis and chemical manipulation of Wnt, BMP, and Hedgehog (Hh) pathways suggest that a common cichlid oral lamina is competent to form teeth or taste buds. Wnt signaling couples tooth and taste bud density and BMP and Hh mediate distinct organ identity. Synthesizing data from fish and mouse, we suggest that the Wnt-BMP-Hh regulatory hierarchy that configures teeth and taste buds on mammalian jaws and tongues may be an evolutionary remnant inherited from ancestors wherein these organs were copatterned from common epithelium.

  14. Whole-Transcriptome Analysis of Differentially Expressed Genes in the Vegetative Buds, Floral Buds and Buds of Chrysanthemum morifolium.

    Directory of Open Access Journals (Sweden)

    Hua Liu

    Full Text Available Chrysanthemum morifolium is an important floral crop that is cultivated worldwide. However, due to a lack of genomic resources, very little information is available concerning the molecular mechanisms of flower development in chrysanthemum.The transcriptomes of chrysanthemum vegetative buds, floral buds and buds were sequenced using Illumina paired-end sequencing technology. A total of 15.4 Gb of reads were assembled into 91,367 unigenes with an average length of 739 bp. A total of 43,137 unigenes showed similarity to known proteins in the Swissprot or NCBI non-redundant protein databases. Additionally, 25,424, 24,321 and 13,704 unigenes were assigned to 56 gene ontology (GO categories, 25 EuKaryotic Orthologous Groups (KOG categories, and 285 Kyoto Encyclopedia of Genes and Genomes (KEGG pathways, respectively. A total of 1,876 differentially expressed genes (DEGs (1,516 up-regulated, 360 down-regulated were identified between vegetative buds and floral buds, and 3,300 DEGs (1,277 up-regulated, 1,706 down-regulated were identified between floral buds and buds. Many genes encoding important transcription factors (e.g., AP2, MYB, MYC, WRKY, NAC and CRT as well as proteins involved in carbohydrate metabolism, protein kinase activity, plant hormone signal transduction, and the defense responses, among others, were considerably up-regulated in floral buds. Genes involved in the photoperiod pathway and flower organ determination were also identified. These genes represent important candidate genes for molecular cloning and functional analysis to study flowering regulation in chrysanthemum.This comparative transcriptome analysis revealed significant differences in gene expression and signaling pathway components between the vegetative buds, floral buds and buds of Chrysanthemum morifolium. A wide range of genes was implicated in regulating the phase transition from vegetative to reproductive growth. These results should aid researchers in the study of

  15. Whole-Transcriptome Analysis of Differentially Expressed Genes in the Vegetative Buds, Floral Buds and Buds of Chrysanthemum morifolium.

    Science.gov (United States)

    Liu, Hua; Sun, Ming; Du, Dongliang; Pan, Huitang; Cheng, Tangren; Wang, Jia; Zhang, Qixiang

    2015-01-01

    Chrysanthemum morifolium is an important floral crop that is cultivated worldwide. However, due to a lack of genomic resources, very little information is available concerning the molecular mechanisms of flower development in chrysanthemum. The transcriptomes of chrysanthemum vegetative buds, floral buds and buds were sequenced using Illumina paired-end sequencing technology. A total of 15.4 Gb of reads were assembled into 91,367 unigenes with an average length of 739 bp. A total of 43,137 unigenes showed similarity to known proteins in the Swissprot or NCBI non-redundant protein databases. Additionally, 25,424, 24,321 and 13,704 unigenes were assigned to 56 gene ontology (GO) categories, 25 EuKaryotic Orthologous Groups (KOG) categories, and 285 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, respectively. A total of 1,876 differentially expressed genes (DEGs) (1,516 up-regulated, 360 down-regulated) were identified between vegetative buds and floral buds, and 3,300 DEGs (1,277 up-regulated, 1,706 down-regulated) were identified between floral buds and buds. Many genes encoding important transcription factors (e.g., AP2, MYB, MYC, WRKY, NAC and CRT) as well as proteins involved in carbohydrate metabolism, protein kinase activity, plant hormone signal transduction, and the defense responses, among others, were considerably up-regulated in floral buds. Genes involved in the photoperiod pathway and flower organ determination were also identified. These genes represent important candidate genes for molecular cloning and functional analysis to study flowering regulation in chrysanthemum. This comparative transcriptome analysis revealed significant differences in gene expression and signaling pathway components between the vegetative buds, floral buds and buds of Chrysanthemum morifolium. A wide range of genes was implicated in regulating the phase transition from vegetative to reproductive growth. These results should aid researchers in the study of flower

  16. HIV Pol inhibits HIV budding and mediates the severe budding defect of Gag-Pol.

    Directory of Open Access Journals (Sweden)

    Xin Gan

    Full Text Available The prevailing hypothesis of HIV budding posits that the viral Gag protein drives budding, and that the Gag p6 peptide plays an essential role by recruiting host-cell budding factors to sites of HIV assembly. HIV also expresses a second Gag protein, p160 Gag-Pol, which lacks p6 and fails to bud from cells, consistent with the prevailing hypothesis of HIV budding. However, we show here that the severe budding defect of Gag-Pol is not caused by the absence of p6, but rather, by the presence of Pol. Specifically, we show that (i the budding defect of Gag-Pol is unaffected by loss of HIV protease activity and is therefore an intrinsic property of the Gag-Pol polyprotein, (ii the N-terminal 433 amino acids of Gag and Gag-Pol are sufficient to drive virus budding even though they lack p6, (iii the severe budding defect of Gag-Pol is caused by a dominant, cis-acting inhibitor of budding in the HIV Pol domain, and (iv Gag-Pol inhibits Gag and virus budding in trans, even at normal levels of Gag and Gag-Pol expression. These and other data support an alternative hypothesis of HIV budding as a process that is mediated by the normal, non-viral pathway of exosome/microvesicle biogenesis.

  17. [Impact of TDZ and NAA on adventitious bud induction and cluster bud multiplication in Tulipa edulis].

    Science.gov (United States)

    Zhu, Li-Fang; Xu, Chao; Zhu, Zai-Biao; Yang, He-Tong; Guo, Qiao-Sheng; Xu, Hong-jian; Ma, Hong-Jian; Zhao, Gui-Hua

    2014-08-01

    To explore the method of explants directly induced bud and establish the tissue culture system of mutiple shoot by means of direct organogenesis, core bud and daughter bulbs (the top of bud stem expanded to form daughter bulb) of T. edulis were used as explants and treated with thidiazuron (TDZ) and 1-naphthlcetic acid (NAA). The results showed that the optimal medium for bud inducted form core bud and daughter bulb were MS + TDZ 2.0 mg x L(-1) + NAA 4.0 mg x L(-1) and MS +TDZ 2.0 mg x L(-1) + NAA 2.0 mg x L(-1) respectively, both of them had a bud induction rate of 72.92%, 79.22%. The optimal medium for cluster buds multiplication was MS + TDZ 0.2 mg x L(-1) + NAA 0.2 mg x L(-1), and proliferation coefficient was 2.23. After proliferation, cluster buds rooting occurred on MS medium with IBA 1.0 mg x L(-1) and the rooting rate was 52.6%, three to five seedlings in each plant. Using core bud and daughter bulb of T. edulis, the optimum medium for adventitious bud directly inducted from daughter bulb, core bud and cluster bud multiplication were screened out and the tissue culture system of multiple shoot by means of direct organogenesis was established.

  18. Repellence of the red bud borer (Resseliella oculiperda) to grafted apple trees by impregnation of budding tape with essential oils

    NARCIS (Netherlands)

    Tol, van R.W.H.M.; Linden, van der A.; Swarts, H.J.; Visser, J.H.

    2007-01-01

    The red bud borer Resseliella oculiperda (Rübs.) is a pest insect of apple trees when rootstocks are grafted with scion buds by shield budding. The female midges are attracted to the wounds of the grafted buds where they lay their eggs. The larvae feed on the cambium and destroy the buds completely

  19. Identification of differentially expressed sequences in bud ...

    African Journals Online (AJOL)

    The suppression subtractive hybridization (SSH) method conducted to generate large-scale expressed sequence tags (EST) was designed to identify gene candidates related to the morphological and physiological differences between the stage before bud differentiation and the stage of bud differentiation of lily. The results ...

  20. Acetylated H4 histone and genomic DNA methylation patterns during bud set and bud burst in Castanea sativa.

    Science.gov (United States)

    Santamaría, Ma Estrella; Hasbún, Rodrigo; Valera, Ma José; Meijón, Mónica; Valledor, Luis; Rodríguez, Jose L; Toorop, Peter E; Cañal, Ma Jesús; Rodríguez, Roberto

    2009-09-01

    The relationships between genomic DNA cytosine methylation, histone H4 acetylation and bud dormancy in Castanea sativa are described. Acetylated H4 histone and genomic DNA methylation patterns showed opposite abundance patterns during bud set and bud burst. Increased and decreased methylation levels in the apical buds coincided with bud set and bud burst, respectively. Intermediate axillary buds were characterized by constant levels of DNA methylation during burst of apical buds and reduced fluctuation in DNA methylation throughout the year, which coincided with the absence of macro-morphological changes. Furthermore, acetylated histone H4 (AcH4) levels from apical buds were higher during bud burst than during bud set, as was demonstrated by immunodetection. Results were validated with three additional C. sativa provenances. Thus, global DNA methylation and AcH4 levels showed opposite patterns and coincided with changes in bud dormancy in C. sativa.

  1. In vitro PROLIFERATION ABILITY OF AXILLARY BUDS IN Musa spp

    African Journals Online (AJOL)

    AISA

    plantain until recently where it was shown with the variety Big Ebanga that axillary buds could equally serve ... As axillary buds have shown mass propagation abilities in Big Ebanga, this explant is tested ... types of buds after four to five sub cultures in all the varieties except for CRBP 39 where the axillary bud exhibits.

  2. Shrinkage of ipsilateral taste buds and hyperplasia of contralateral taste buds following chorda tympani nerve transection

    Directory of Open Access Journals (Sweden)

    Yi-ke Li

    2015-01-01

    Full Text Available The morphological changes that occur in the taste buds after denervation are not well understood in rats, especially in the contralateral tongue epithelium. In this study, we investigated the time course of morphological changes in the taste buds following unilateral nerve transection. The role of the trigeminal component of the lingual nerve in maintaining the structural integrity of the taste buds was also examined. Twenty-four Sprague-Dawley rats were randomly divided into three groups: control, unilateral chorda tympani nerve transection and unilateral chorda tympani nerve transection + lingual nerve transection. Rats were allowed up to 42 days of recovery before being euthanized. The taste buds were visualized using a cytokeratin 8 antibody. Taste bud counts, volumes and taste receptor cell numbers were quantified and compared among groups. No significant difference was detected between the chorda tympani nerve transection and chorda tympani nerve transection + lingual nerve transection groups. Taste bud counts, volumes and taste receptor cell numbers on the ipsilateral side all decreased significantly compared with control. On the contralateral side, the number of taste buds remained unchanged over time, but they were larger, and taste receptor cells were more numerous postoperatively. There was no evidence for a role of the trigeminal branch of the lingual nerve in maintaining the structural integrity of the anterior taste buds.

  3. Complex bud architecture and cell-specific chemical patterns enable supercooling of Picea abies bud primordial

    Science.gov (United States)

    Bud primordia of Picea abies, despite a frozen shoot, stay ice free down to -50 °C by a mechanism termed supercooling whose biophysical and biochemical requirements are poorly understood. Bud architecture was assessed by 3D-reconstruction, supercooling and freezing patterns by infrared video thermog...

  4. Taste buds as peripheral chemosensory processors.

    Science.gov (United States)

    Roper, Stephen D

    2013-01-01

    Taste buds are peripheral chemosensory organs situated in the oral cavity. Each taste bud consists of a community of 50-100 cells that interact synaptically during gustatory stimulation. At least three distinct cell types are found in mammalian taste buds - Type I cells, Receptor (Type II) cells, and Presynaptic (Type III) cells. Type I cells appear to be glial-like cells. Receptor cells express G protein-coupled taste receptors for sweet, bitter, or umami compounds. Presynaptic cells transduce acid stimuli (sour taste). Cells that sense salt (NaCl) taste have not yet been confidently identified in terms of these cell types. During gustatory stimulation, taste bud cells secrete synaptic, autocrine, and paracrine transmitters. These transmitters include ATP, acetylcholine (ACh), serotonin (5-HT), norepinephrine (NE), and GABA. Glutamate is an efferent transmitter that stimulates Presynaptic cells to release 5-HT. This chapter discusses these transmitters, which cells release them, the postsynaptic targets for the transmitters, and how cell-cell communication shapes taste bud signaling via these transmitters. Copyright © 2012 Elsevier Ltd. All rights reserved.

  5. Induction of adventitious buds on buds of Norway spruce (picea abies) grown in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Von Arnold, S.; Eriksson, T.

    1979-01-01

    Isolated resting vegetative buds of P. abies were cultured on a defined medium to which auxins or cytokinins were added. At concentrations of 10 to the power of -6 to 10 to the power of -4 M, 6-benzylaminopurine (BAP) or 6-(3-methyl-2-buten-1-ylamino)-purine (2iP) induced formation of adentitious bud primordia; these arose from meristems formed de novo in the needle primordia. Adventitious buds developed when these buds were transferred to medium lacking cytokinin. At lower concentrations of BAP or 2iP, only elongation of needle primordia was induced; at higher concentrations, buds were killed. Other growth regulators induced only needle primordia elongation and were also lethal at higher concentrations. No major differences were found in the ability to form adventitious buds among buds from: a 75-year-old hedge, 5-year-old greenhouse-grown plants, and trees in a stand near Uppsala, Sweden ranging in age from 5 to 50 year old.

  6. Activation of the retroviral budding factor ALIX.

    Science.gov (United States)

    Zhai, Qianting; Landesman, Michael B; Chung, Hyo-Young; Dierkers, Adam; Jeffries, Cy M; Trewhella, Jill; Hill, Christopher P; Sundquist, Wesley I

    2011-09-01

    The cellular ALIX protein functions within the ESCRT pathway to facilitate intralumenal endosomal vesicle formation, the abscission stage of cytokinesis, and enveloped virus budding. Here, we report that the C-terminal proline-rich region (PRR) of ALIX folds back against the upstream domains and auto-inhibits V domain binding to viral late domains. Mutations designed to destabilize the closed conformation of the V domain opened the V domain, increased ALIX membrane association, and enhanced virus budding. These observations support a model in which ALIX activation requires dissociation of the autoinhibitory PRR and opening of the V domain arms.

  7. Identification and Quality Assessment of Chrysanthemum Buds by CE Fingerprinting

    Directory of Open Access Journals (Sweden)

    Xiaoping Xing

    2015-01-01

    Full Text Available A simple and efficient fingerprinting method for chrysanthemum buds was developed with the aim of establishing a quality control protocol based on biochemical makeup. Chrysanthemum bud samples were successively extracted by water and alcohol. The fingerprints of the chrysanthemum buds samples were obtained using capillary electrophoresis and electrochemical detection (CE-ED employing copper and carbon working electrodes to capture all of the chemical information. 10 batches of chrysanthemum buds were collected from different regions and various factories to establish the baseline fingerprint. The experimental data of 10 batches electropherogram buds by CE were analyzed by correlation coefficient and the included angle cosine methods. A standard chrysanthemum bud fingerprint including 24 common peaks was established, 12 from each electrode, which was successfully applied to identify and distinguish between chrysanthemum buds from 2 other chrysanthemum species. These results demonstrate that fingerprint analysis can be used as an important criterion for chrysanthemum buds quality control.

  8. Total RNA quality of lyophilized and cryopreserved dormant grapevine buds

    Directory of Open Access Journals (Sweden)

    Claudia Vanessa García-Baldenegro

    2015-03-01

    Conclusion: High-quality RNA that is useful for downstream applications was obtained from freeze-dried dormant grapevine bud tissue, similarly to the RNA obtained from cryopreserved dormant grapevine buds.

  9. Tumor budding in colorectal carcinoma assessed by cytokeratin immunostaining and budding areas: possible involvement of c-Met.

    Science.gov (United States)

    Satoh, Keisuke; Nimura, Satoshi; Aoki, Mikiko; Hamasaki, Makoto; Koga, Kaori; Iwasaki, Hiroshi; Yamashita, Yuichi; Kataoka, Hiroaki; Nabeshima, Kazuki

    2014-11-01

    Tumor budding/sprouting has been shown to be an independent adverse prognostic factor in T1 and T3N0 colorectal carcinomas, however, its assessment could be improved by more accurate identification of budding carcinoma cells and consideration of budding areas. Moreover, tumor budding mechanisms are yet to be defined. In this study, we evaluated the identification of budding tumor cells by either H&E staining alone or H&E with immunohistochemistry and developed a scoring system based on budding grades and areas. We examined whether the budding score correlated with clinicopathologic features and prognosis and the association between tumor budding/sprouting and c-Met protein expression and phosphorylation and MET gene copy numbers because c-Met is known to play an important role in colorectal carcinoma tumorigenesis. Cytokeratin immunohistochemistry could identify tumors with shorter disease-free survival (DFS) from the low-grade budding group assessed with H&E alone. High budding scores based on budding grade and area were more significantly correlated with DFS than scores obtained using the budding grade alone. In tumors with a high budding score, c-Met expression and phosphorylation levels and MET gene copy numbers were significantly increased at the invasive front compared with those in superficial tumor portions. This study showed for the first time that high levels of phospho-c-Met at the invasive front were significantly associated with a high budding score and shorter DFS. In conclusion, a budding score assessed by budding grades and budding-positive areas correlates highly with clinicopathologic aggressive features of colorectal carcinoma. © 2014 The Authors. Cancer Science published by Wiley Publishing Asia Pty Ltd on behalf of Japanese Cancer Association.

  10. Sprouting of dormant buds on border trees

    Science.gov (United States)

    G.R., Jr. Trimble; H. Clay Smith; H. Clay Smith

    1970-01-01

    As part of an evaluation of silvicultura1 systems used in managing Appalachian hardwoods, we are studying degrade of border trees surrounding harvest-cut openings made in the patch cutting and group selection systems. One facet of this research dealt with determining what portion of visually evident dormant buds on border tree boles sprouted when the openings were cut...

  11. Bipolar budding in yeasts - an electron microscope study

    NARCIS (Netherlands)

    Kreger-van Rij, N.J.W.; Veenhuis, M.

    1971-01-01

    Bud formation in yeasts with bipolar budding was studied by electron microscopy of thin sections. Budding in yeasts of the species Saccharomycodes ludwigii, Hanseniaspora valbyensis and Wickerhamia fluorescens resulted in concentric rings of scar ridges on the wall of the mother cell. The wall

  12. Overhead irrigation increased winter chilling and floral bud ...

    African Journals Online (AJOL)

    Eucalyptus nitens requires a sufficiently cold winter to produce flower buds. In areas in South Africa where E. nitens commercial plantations as well as breeding and production seed orchards are located, winter chilling is often insufficient for floral bud initiation. Hence, under such conditions, E. nitens floral bud and seed ...

  13. Taste buds in the palatal mucosa of snakes | Berkhoudt | African ...

    African Journals Online (AJOL)

    An examination of the oral mucosa of Crotalus and several Scolecophidia revealed the presence of taste buds. The taste buds in these two divergent groups of snakes are similar in appearance, and correspond to previous descriptions of gustatory organs in other reptiles. Few taste buds were present in any specimen, and ...

  14. Effect of temperature on development and growth potential of axillary buds in roses

    NARCIS (Netherlands)

    Marcelis-van Acker, C.A.M.

    1995-01-01

    The effect of temperature during axillary bud formation on axillary bud development and subsequent shoot growth was investigated. Growth potential of the axillary buds was studied either in situ, by pruning the parent shoot above the bud, or in isolation, by grafting the bud or by culturing the bud

  15. Ontogeny of axillary buds and shoots in roses: Leaf initiation and pith development.

    NARCIS (Netherlands)

    Marcelis-van Acker, C.A.M.

    1994-01-01

    The ontogeny of an axillary bud (in the middle region of a shoot) from initiation up to flowering of the subsequent shoot was studied. The first secondary buds appeared in the axillary bud (primary bud) when the leaf subtending the primary bud unfolded. By that time, the primary bud contained seven

  16. Epithelial mesenchymal transition and tumor budding in aggressive colorectal cancer: Tumor budding as oncotarget

    Science.gov (United States)

    Zlobec, Inti; Lugli, Alessandro

    2010-01-01

    Epithelial mesenchymal transition (EMT) is proposed as a critical mechanism for the acquisition of malignant phenotypes by epithelial cells. In colorectal cancer, tumor cells having undergone EMT are histologically represented by the presence of tumor buds defined as single cells or small clusters of de-differentiated tumor cells at the invasive front. Tumor budding is not a static, histological feature rather it represents a snap-shot of a dynamic process undertaken by an aggressive tumor with the potential to disseminate and metastasize. Strong, consistent evidence shows that tumor budding is a predictor of lymph node metastasis, distant metastatic disease, local recurrence, worse overall and disease-free survival time and an independent prognostic factor. Moreover, the International Union against Cancer (UICC) recognizes tumor budding as a highly relevant, additional prognostic parameter. The aim of this review is to summarize the evidence supporting the implementation of tumor budding into diagnostic pathology and patient management and additionally to illustrate its worthiness as a potential therapeutic target. PMID:21317460

  17. Tumor budding in colorectal cancer--ready for diagnostic practice?

    Science.gov (United States)

    Koelzer, Viktor H; Zlobec, Inti; Lugli, Alessandro

    2016-01-01

    Tumor budding is an important additional prognostic factor for patients with colorectal cancer (CRC). Defined as the presence of single tumor cells or small clusters of up to 5 cells in the tumor stroma, tumor budding has been likened to an epithelial-mesenchymal transition. Based on well-designed retrospective studies, tumor budding is linked to adverse outcome of CRC patients in 3 clinical scenarios: (1) in malignant polyps, detection of tumor buds is a risk factor for lymph node metastasis indicating the need for colorectal surgery; (2) tumor budding in stage II CRC is a highly adverse prognostic indicator and may aid patient selection for adjuvant therapy; (3) in the preoperative setting, presence of tumor budding in biopsy material may help to identify high-risk rectal cancer patients for neoadjuvant therapy. However, lack of consensus guidelines for standardized assessment still limits reporting in daily diagnostic practice. This article provides a practical and comprehensive overview on tumor budding aimed at the practicing pathologist. First, we review the prognostic value of tumor budding for the management of colon and rectal cancer patients. Second, we outline a practical, evidence-based proposal for the assessment of tumor budding in the daily sign-out. Last, we summarize the current knowledge of the molecular characteristics of high-grade budding tumors in the context of personalized treatment approaches and biomarker discovery. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Development Correlations of the Buds of Grapevine (Vitis vinifera L.

    Directory of Open Access Journals (Sweden)

    Liliana ROTARU

    2010-06-01

    Full Text Available The development characteristics of different buds of the grapevine are mainly related by stimulation and/or inhibition effects, the action of which is still inexplicable. The present study examines the development dynamics of the buds of a one-year old branch after excision of different buds and the application of ?-naphtyl acetic acid (ANA, as well as the growth capacity of each bud individually. We verified the effects of acrotony cited previously by various researchers. These effects are due to different developmental characteristics of which could to lay the groundwork for the improvement of different productions methods.

  19. Essential Oil of Betula pendula Roth. Buds

    Directory of Open Access Journals (Sweden)

    Betül Demirci

    2004-01-01

    Full Text Available The essential oil of Betula pendula Roth. buds was obtained using both hydrodistillation and microdistillation techniques and their chemical compositions were analyzed using both gas chromatography (GC and gas chromatography–mass spectrometry (GC-MS. Overall, more than 50 compounds were identified representing 80% and 92% for hydrodistillation and microdistillation, respectively. The main components (by hydrodistillation and microdistillation, respectively found were α-copaene (12% and 10%, germacrene D (11% and 18% and δ-cadinene (11% and 15% in the analyzed essential oils. The microdistillation technique proved to be a useful tool and compliant alternative when compared to hydrodistillation.

  20. Recommendations for reporting tumor budding in colorectal cancer based on the International Tumor Budding Consensus Conference (ITBCC) 2016

    DEFF Research Database (Denmark)

    Lugli, Alessandro; Kirsch, Richard; Ajioka, Yoichi

    2017-01-01

    Tumor budding is a well-established independent prognostic factor in colorectal cancer but a standardized method for its assessment has been lacking. The primary aim of the International Tumor Budding Consensus Conference (ITBCC) was to reach agreement on an international, evidence......-based standardized scoring system for tumor budding in colorectal cancer. The ITBCC included nine sessions with presentations, a pre-meeting survey and an e-book covering the key publications on tumor budding in colorectal cancer. The Grading of Recommendation Assessment, Development and Evaluation' method was used...... to determine the strength of recommendations and quality of evidence. The following 10 statements achieved consensus: Tumor budding is defined as a single tumor cell or a cell cluster consisting of four tumor cells or less (22/22, 100%). Tumor budding is an independent predictor of lymph node metastases in pT1...

  1. Complex bud architecture and cell-specific chemical patterns enable supercooling of Picea abies bud primordia.

    Science.gov (United States)

    Kuprian, Edith; Munkler, Caspar; Resnyak, Anna; Zimmermann, Sonja; Tuong, Tan D; Gierlinger, Notburga; Müller, Thomas; Livingston, David P; Neuner, Gilbert

    2017-12-01

    Bud primordia of Picea abies, despite a frozen shoot, stay ice free down to -50 °C by a mechanism termed supercooling whose biophysical and biochemical requirements are poorly understood. Bud architecture was assessed by 3D-reconstruction, supercooling and freezing patterns by infrared video thermography, freeze dehydration and extraorgan freezing by water potential measurements, and cell-specific chemical patterns by Raman microscopy and mass spectrometry imaging. A bowl-like ice barrier tissue insulates primordia from entrance by intrinsic ice. Water repellent and densely packed bud scales prevent extrinsic ice penetration. At -18 °C, break-down of supercooling was triggered by intrinsic ice nucleators whereas the ice barrier remained active. Temperature-dependent freeze dehydration (-0.1 MPa K-1 ) caused accumulation of extraorgan ice masses that by rupture of the shoot, pith tissue are accommodated in large voids. The barrier tissue has exceptionally pectin-rich cell walls and intercellular spaces, and the cell lumina were lined or filled with proteins, especially near the primordium. Primordial cells close to the barrier accumulate di, tri and tetrasaccharides. Bud architecture efficiently prevents ice penetration, but ice nucleators become active inside the primordium below a temperature threshold. Biochemical patterns indicate a complex cellular interplay enabling supercooling and the necessity for cell-specific biochemical analysis. © 2017 The Authors Plant, Cell & Environment Published by John Wiley & Sons Ltd.

  2. Electrochemical regulation of budding yeast polarity.

    Directory of Open Access Journals (Sweden)

    Armin Haupt

    2014-12-01

    Full Text Available Cells are naturally surrounded by organized electrical signals in the form of local ion fluxes, membrane potential, and electric fields (EFs at their surface. Although the contribution of electrochemical elements to cell polarity and migration is beginning to be appreciated, underlying mechanisms are not known. Here we show that an exogenous EF can orient cell polarization in budding yeast (Saccharomyces cerevisiae cells, directing the growth of mating projections towards sites of hyperpolarized membrane potential, while directing bud emergence in the opposite direction, towards sites of depolarized potential. Using an optogenetic approach, we demonstrate that a local change in membrane potential triggered by light is sufficient to direct cell polarization. Screens for mutants with altered EF responses identify genes involved in transducing electrochemical signals to the polarity machinery. Membrane potential, which is regulated by the potassium transporter Trk1p, is required for polarity orientation during mating and EF response. Membrane potential may regulate membrane charges through negatively charged phosphatidylserines (PSs, which act to position the Cdc42p-based polarity machinery. These studies thus define an electrochemical pathway that directs the orientation of cell polarization.

  3. Electrochemical Regulation of Budding Yeast Polarity

    Science.gov (United States)

    Piel, Matthieu; Chang, Fred; Minc, Nicolas

    2014-01-01

    Cells are naturally surrounded by organized electrical signals in the form of local ion fluxes, membrane potential, and electric fields (EFs) at their surface. Although the contribution of electrochemical elements to cell polarity and migration is beginning to be appreciated, underlying mechanisms are not known. Here we show that an exogenous EF can orient cell polarization in budding yeast (Saccharomyces cerevisiae) cells, directing the growth of mating projections towards sites of hyperpolarized membrane potential, while directing bud emergence in the opposite direction, towards sites of depolarized potential. Using an optogenetic approach, we demonstrate that a local change in membrane potential triggered by light is sufficient to direct cell polarization. Screens for mutants with altered EF responses identify genes involved in transducing electrochemical signals to the polarity machinery. Membrane potential, which is regulated by the potassium transporter Trk1p, is required for polarity orientation during mating and EF response. Membrane potential may regulate membrane charges through negatively charged phosphatidylserines (PSs), which act to position the Cdc42p-based polarity machinery. These studies thus define an electrochemical pathway that directs the orientation of cell polarization. PMID:25548923

  4. Tumor budding in upper gastrointestinal carcinomas

    Directory of Open Access Journals (Sweden)

    Viktor Hendrik Koelzer

    2014-08-01

    Full Text Available The basis of personalized medicine in oncology is the prediction of an individual’s risk of relapse and death from disease. The presence of tumor budding (TB at the tumor-host interface of gastrointestinal cancers has been recognized as a hallmark of unfavorable disease biology. TB is defined as the presence of dedifferentiated cells or small clusters of up to five cells at the tumor invasive front and can be observed in aggressive carcinomas of the esophagus, stomach, pancreas, ampulla, colon and rectum. Presence of TB reproducibly correlates with advanced tumor stage, frequent lymphovascular invasion, nodal and distant metastasis. The UICC has officially recognized TB as additional independent prognostic factor in cancers of the colon and rectum. Recent studies have also characterized TB as a promising prognostic indicator for clinical management of esophageal squamous cell carcinoma, adenocarcinoma of the gastro-esophageal junction and gastric adenocarcinoma. However, several important issues have to be addressed for application in daily diagnostic practice: 1 Validation of prognostic scoring systems for tumor budding in large, multi-center studies 2 Consensus on the optimal assessment method 3 Inter-observer reproducibility. This review provides a comprehensive analysis of TB in cancers of the upper gastrointestinal tract including critical appraisal of perspectives for further study.

  5. Field and laboratory investigations of budding in the tetillid sponge Cinachyrella cavernosa

    Digital Repository Service at National Institute of Oceanography (India)

    Singh, A.; Thakur, N.L.

    regression analysis of physico-chemical factors revealed temperature as the most prominent factor regulating the intensity of budding. Based on size and morphology, three stages of sponge buds were defined. The production of buds was found to be asynchronous...

  6. Recommendations for reporting tumor budding in colorectal cancer based on the International Tumor Budding Consensus Conference (ITBCC) 2016.

    Science.gov (United States)

    Lugli, Alessandro; Kirsch, Richard; Ajioka, Yoichi; Bosman, Fred; Cathomas, Gieri; Dawson, Heather; El Zimaity, Hala; Fléjou, Jean-François; Hansen, Tine Plato; Hartmann, Arndt; Kakar, Sanjay; Langner, Cord; Nagtegaal, Iris; Puppa, Giacomo; Riddell, Robert; Ristimäki, Ari; Sheahan, Kieran; Smyrk, Thomas; Sugihara, Kenichi; Terris, Benoît; Ueno, Hideki; Vieth, Michael; Zlobec, Inti; Quirke, Phil

    2017-09-01

    Tumor budding is a well-established independent prognostic factor in colorectal cancer but a standardized method for its assessment has been lacking. The primary aim of the International Tumor Budding Consensus Conference (ITBCC) was to reach agreement on an international, evidence-based standardized scoring system for tumor budding in colorectal cancer. The ITBCC included nine sessions with presentations, a pre-meeting survey and an e-book covering the key publications on tumor budding in colorectal cancer. The 'Grading of Recommendation Assessment, Development and Evaluation' method was used to determine the strength of recommendations and quality of evidence. The following 10 statements achieved consensus: tumor budding is defined as a single tumor cell or a cell cluster consisting of four tumor cells or less (22/22, 100%). Tumor budding is an independent predictor of lymph node metastases in pT1 colorectal cancer (23/23, 100%). Tumor budding is an independent predictor of survival in stage II colorectal cancer (23/23, 100%). Tumor budding should be taken into account along with other clinicopathological features in a multidisciplinary setting (23/23, 100%). Tumor budding is counted on H&E (19/22, 86%). Intratumoral budding exists in colorectal cancer and has been shown to be related to lymph node metastasis (22/22, 100%). Tumor budding is assessed in one hotspot (in a field measuring 0.785 mm(2)) at the invasive front (22/22, 100%). A three-tier system should be used along with the budding count in order to facilitate risk stratification in colorectal cancer (23/23, 100%). Tumor budding and tumor grade are not the same (23/23, 100%). Tumor budding should be included in guidelines/protocols for colorectal cancer reporting (23/23, 100%). Members of the ITBCC were able to reach strong consensus on a single international, evidence-based method for tumor budding assessment and reporting. It is proposed that this method be incorporated into colorectal cancer

  7. Bud initiation and optimum harvest date in Brussels sprouts

    NARCIS (Netherlands)

    Everaarts, A.P.; Sukkel, W.

    1999-01-01

    For six cultivars of Brussels sprouts (Brassica oleracea var. gemmifera) with a decreasing degree of earliness, or optimum harvest date, the time of bud initiation was determined during two seasons. Fifty percent of the plants had initiated buds between 60 and 75 days after planting (DAP) in 1994

  8. Cell to cell signalling during vertebrate limb bud development

    NARCIS (Netherlands)

    Panman, Lia

    2004-01-01

    Communication between cells is essential during embryonic development. The vertebrate limb bud provides us a model to study signalling interactions between cells during patterning of embryonic tissues and organogenesis. In chapter 1 I give an introduction about limb bud development that is focussed

  9. Floral bud distortion in soybean and incidence in Central India ...

    African Journals Online (AJOL)

    Floral bud distortion in soybean and incidence in Central India. V Jadhav Pravin, SS Mane, RS Nandanwar, PB Kale, MS Dudhare, MP Moharil, RG Dani. Abstract. We describe a peculiar and often harmful budding disorder in soybean, leading to huge yield loss in India. To determine the prevalence of floral distortion in ...

  10. Photocontrol of bud burst involves gibberellin biosynthesis in Rosa sp.

    Science.gov (United States)

    Choubane, Djillali; Rabot, Amélie; Mortreau, Eric; Legourrierec, Jose; Péron, Thomas; Foucher, Fabrice; Ahcène, Youyou; Pelleschi-Travier, Sandrine; Leduc, Nathalie; Hamama, Latifa; Sakr, Soulaiman

    2012-09-01

    Light is a critical determinant of plant shape by controlling branching patterns and bud burst in many species. To gain insight into how light induces bud burst, we investigated whether its inductive effect in rose was related to gibberellin (GA) biosynthesis. In axillary buds of beheaded plants subject to light, the expression of two GA biosynthesis genes (RoGA20ox and RoGA3ox) was promptly and strongly induced, while that of a GA-catabolism genes (RoGA2ox) was reduced. By contrast, lower expression levels of these two GA biosynthesis genes were found in darkness, and correlated with a total inhibition of bud burst. This effect was dependent on both light intensity and quality. In in vitro cultured buds, the inductive effect of light on the growth of preformed leaves and SAM organogenic activity was inhibited by ancymidol and paclobutrazol, two effectors of GA biosynthesis. This effect was concentration-dependent, and negated by GA(3). However, GA(3) alone could not rescue bud burst in the dark. GA biosynthesis was also required for the expression and activity of a vacuolar invertase, and therefore for light-induced sugar metabolism within buds. These findings are evidence that GA biosynthesis contributes to the light effect on bud burst and lay the foundations of a better understanding of its exact role in plant branching. Copyright © 2012 Elsevier GmbH. All rights reserved.

  11. An elastic model of partial budding of retroviruses

    Science.gov (United States)

    Zhang, Rui; Nguyen, Toan

    2008-03-01

    Retroviruses are characterized by their unique infection strategy of reverse transcription, in which the genetic information flows from RNA back to DNA. The most well known representative is the human immunodeficiency virus (HIV). Unlike budding of traditional enveloped viruses, retrovirus budding happens together with the formation of spherical virus capsids at the cell membrane. Led by this unique budding mechanism, we proposed an elastic model of retrovirus budding in this work. We found that if the lipid molecules of the membrane are supplied fast enough from the cell interior, the budding always proceeds to completion. In the opposite limit, there is an optimal size of partially budded virions. The zenith angle of these partially spherical capsids, α, is given by α˜(2̂/κσ)^1/4, where κ is the bending modulus of the membrane, σ is the surface tension of the membrane, and τ characterizes the strength of capsid protein interaction. If τ is large enough such that α˜π, the budding is complete. Our model explained many features of retrovirus partial budding observed in experiments.

  12. The prognostic value of tumor budding in invasive breast cancer.

    Science.gov (United States)

    Liang, Fenli; Cao, Wei; Wang, Yili; Li, Linrui; Zhang, Guanjun; Wang, Zhuo

    2013-05-01

    We investigated the prognostic value of tumor budding in 160 cases of operable invasive ductal carcinoma, not otherwise specified (IDC-NOS). The number of buds was counted in H&E slides with a maximal invasive margin in a 0.950mm(2) field of vision (200×). According to a cut-off score selected by ROC analysis, the cohort was dichotomized into a low (0-7 budding foci, 107 cases, 66.9%) and a high-grade budding group (8 or more budding foci, 53 cases, 33.1%). The inter-observer test showed a good reproducibility with 0.717 as the К value. High-grade budding was significantly associated with the presence of lymphovascular invasion (LVI) (P=0.001), larger tumor size (P=0.014), and worse clinical outcome (Pbudded cells at the margin displayed epithelial mesenchymal transition (EMT)-like molecular phenotype and decreased proliferative activity. Survival analyses revealed that tumor budding (HR 4.275, Ptumor size (HR 2.468, P=0.002), node status (HR 2.362, Ptumor budding is a reproducible, significant, and independent histopathological prognostic factor in IDC-NOS. Copyright © 2013 Elsevier GmbH. All rights reserved.

  13. Impact of peritumoral and intratumoral budding in esophageal adenocarcinomas.

    Science.gov (United States)

    Thies, Svenja; Guldener, Lars; Slotta-Huspenina, Julia; Zlobec, Inti; Koelzer, Viktor H; Lugli, Alessandro; Kröll, Dino; Seiler, Christian A; Feith, Marcus; Langer, Rupert

    2016-06-01

    Tumor budding has prognostic significance in many carcinomas and is defined as the presence of detached isolated single cells or small cell clusters up to 5 cells at the invasion front (peritumoral budding [PTB]) or within the tumor (intratumoral budding [ITB]). For esophageal adenocarcinomas (EACs), there are currently only few data about the impact of this morphological feature. We investigated tumor budding in a large collective of 200 primarily resected EACs. Pancytokeratin staining was demonstrated to be superior to hematoxylin and eosin staining for the detection of buds with substantial to excellent interobserver agreement and used for subsequent analysis. PTB and ITB were scored across 10 high-power fields (HPFs). The median count of tumor buds was 130/10 HPFs for PTB (range, 2-593) and 80/10 HPFs for ITB (range, 1-656). PTB and ITB correlated significantly with each other (r = 0.9; P tumor categories (P tumors with lymph node metastases (P tumor grading (P budding is associated with aggressive tumor phenotype. Assessment of tumor budding, especially ITB, may provide additional prognostic information about tumor behavior and may be useful in specific cases for risk stratification of EAC patients. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Plantlets from encapsulated shoot buds of Catalpa ovata G. Don

    Directory of Open Access Journals (Sweden)

    Halina Wysokińska

    2014-01-01

    Full Text Available Shoot buds isolated from in vitro shoot cultures of Catalpa ovata G. Don were encapsulated using 3% sodium alginate with sucrose (3% and 50 mM calcium chloride. The morphogenic response of encapsulated buds was affected by such factors, like composition of the media and the presence of growth regulators. The highest frequency of plantlet germination from encapsulated buds (70% within 4 weeks was obtained on Woody Plant medium (WP (Lloyd and McCown 1980 containing indole-3-butyric acid (IBA (1 mg/l. The process was substantially inhibited by cold-storage (4oC of encapsulated buds. In this case, the frequency response ranged from 3% to 22% dependent on storage period (28 or 42 days and the presence of the paraffin coat covering the alginate capsules. The plantlets developed from both unstored and stored encapsulated buds of C. ovata were transplanted to soil and grew in pots to phenotypically normal plants.

  15. Cytokinins and polar transport of auxin in axillary pea buds

    Directory of Open Access Journals (Sweden)

    Petr Kalousek

    2010-01-01

    Full Text Available The influence of cytokinin on auxin transport during release of axillary buds from apical dominance was studied. Expression of auxin-carrier coding genes PsAUX1 (AUXIN RESISTANT 1 and PsPIN1 (PIN-FORMED 1 was explored in axillary buds of the 2nd node of 7-day pea plants (Pisum sativum L. cv. Vladan after decapitation or after exogenous application of benzyladenine (6-benzylaminopurine onto axillary buds of intact plants. Localization of the PsPIN1 protein, the key factor for polar transport of auxin in axillary buds, was visualised by immunohistochemistry. After exogenous application of cytokinin the expression of PsAUX1 and PsPIN1 rapidly increased with a simultaneous rapid decrease in PsDRM1 and PsAD1 expression – genes related to bud dormancy. The same changes in expression were observed after decapitation, however they were markedly slower. The PsPIN1 auxin efflux carrier in the inhibited axillary buds of intact plants was localised in a non-polar manner. After exogenous application of cytokinin gradual polarisation of the PsPIN1 protein occurred on the basal pole of polar auxin transport competent cells. Despite the fact that direct auxin application to buds of intact plants led to an increase in PsAUX1 and PsPIN1 expression, the buds remained dormant (non-growing what was accompanied by persistent expression of the dormancy markers PsDRM1 and PsAD1. The results indicate a possible effect of cytokinins on biosynthesis, and/or transport of auxin in axillary buds and they highlight the importance of auxin-cytokinin crosstalk in the regulation of bud outgrowth after breaking of apical dominance.

  16. High tumor budding stratifies breast cancer with metastatic properties.

    Science.gov (United States)

    Salhia, Bodour; Trippel, Mafalda; Pfaltz, Katrin; Cihoric, Nikola; Grogg, André; Lädrach, Claudia; Zlobec, Inti; Tapia, Coya

    2015-04-01

    Tumor budding refers to single or small cluster of tumor cells detached from the main tumor mass. In colon cancer high tumor budding is associated with positive lymph nodes and worse prognosis. Therefore, we investigated the value of tumor budding as a predictive feature of lymph node status in breast cancer (BC). Whole tissue sections from 148 surgical resection specimens (SRS) and 99 matched preoperative core biopsies (CB) with invasive BC of no special type were analyzed on one slide stained with pan-cytokeratin. In SRS, the total number of intratumoral (ITB) and peripheral tumor buds (PTB) in ten high-power fields (HPF) were counted. A bud was defined as a single tumor cell or a cluster of up to five tumor cells. High tumor budding equated to scores averaging >4 tumor buds across 10HPFs. In CB high tumor budding was defined as ≥10 buds/HPF. The results were correlated with pathological parameters. In SRS high PTB stratified BC with lymph node metastases (p ≤ 0.03) and lymphatic invasion (p ≤ 0.015). In CB high tumor budding was significantly (p = 0.0063) associated with venous invasion. Pathologists are able, based on morphology, to categorize BC into a high and low risk groups based in part on lymph node status. This risk assessment can be easily performed during routine diagnostics and it is time and cost effective. These results suggest that high PTB is associated with loco-regional metastasis, highlighting the possibility that this tumor feature may help in therapeutic decision-making.

  17. Budding yeast for budding geneticists: a primer on the Saccharomyces cerevisiae model system.

    Science.gov (United States)

    Duina, Andrea A; Miller, Mary E; Keeney, Jill B

    2014-05-01

    The budding yeast Saccharomyces cerevisiae is a powerful model organism for studying fundamental aspects of eukaryotic cell biology. This Primer article presents a brief historical perspective on the emergence of this organism as a premier experimental system over the course of the past century. An overview of the central features of the S. cerevisiae genome, including the nature of its genetic elements and general organization, is also provided. Some of the most common experimental tools and resources available to yeast geneticists are presented in a way designed to engage and challenge undergraduate and graduate students eager to learn more about the experimental amenability of budding yeast. Finally, a discussion of several major discoveries derived from yeast studies highlights the far-reaching impact that the yeast system has had and will continue to have on our understanding of a variety of cellular processes relevant to all eukaryotes, including humans.

  18. Cell polarization in budding and fission yeasts.

    Science.gov (United States)

    Martin, Sophie G; Arkowitz, Robert A

    2014-03-01

    Polarization is a fundamental cellular property, which is essential for the function of numerous cell types. Over the past three to four decades, research using the best-established yeast systems in cell biological research, Saccharomyces cerevisiae (or budding yeast) and Schizosaccharomyces pombe (or fission yeast), has brought to light fundamental principles governing the establishment and maintenance of a polarized, asymmetric state. These two organisms, though both ascomycetes, are evolutionarily very distant and exhibit distinct shapes and modes of growth. In this review, we compare and contrast the two systems. We first highlight common cell polarization pathways, detailing the contribution of Rho GTPases, the cytoskeleton, membrane trafficking, lipids, and protein scaffolds. We then contrast the major differences between the two organisms, describing their distinct strategies in growth site selection and growth zone dimensions and compartmentalization, which may be the basis for their distinct shapes. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  19. Analyzing DNA replication checkpoint in budding yeast.

    Science.gov (United States)

    Hustedt, Nicole; Shimada, Kenji

    2014-01-01

    Checkpoints are conserved mechanisms that prevent progression into the next phase of the cell cycle when cells are unable to accomplish the previous event properly. Cells also possess a surveillance mechanism called the DNA replication checkpoint, which consists of a conserved kinase cascade that is provoked by insults that block or slow down replication fork progression. In the budding yeast Saccharomyces cerevisiae, the DNA replication checkpoint controls the timing of S-phase events such as origin firing and spindle elongation. This checkpoint also upregulates dNTP pools and maintains the replication fork structure in order to resume DNA replication after replication block. Many replication checkpoint factors have been found to be tumor suppressors, highlighting the importance of this checkpoint pathway in human health. Here we describe a series of protocols to analyze the DNA replication checkpoint in S. cerevisiae.

  20. Decellularized Tooth Bud Scaffolds for Tooth Regeneration.

    Science.gov (United States)

    Zhang, W; Vazquez, B; Oreadi, D; Yelick, P C

    2017-05-01

    Whole tooth regeneration approaches currently are limited by our inability to bioengineer full-sized, living replacement teeth. Recently, decellularized organ scaffolds have shown promise for applications in regenerative medicine by providing a natural extracellular matrix environment that promotes cell attachment and tissue-specific differentiation leading to full-sized organ regeneration. We hypothesize that decellularized tooth buds (dTBs) created from unerupted porcine tooth buds (TBs) can be used to guide reseeded dental cell differentiation to form whole bioengineered teeth, thereby providing a potential off-the-shelf scaffold for whole tooth regeneration. Porcine TBs were harvested from discarded 6-mo-old pig jaws, and decellularized by successive sodium dodecyl sulfate/Triton-X cycles. Four types of replicate implants were used in this study: 1) acellular dTBs; 2) recellularized dTBs seeded with porcine dental epithelial cells, human dental pulp cells, and human umbilical vein endothelial cells (recell-dTBs); 3) dTBs seeded with bone morphogenetic protein (BMP)-2 (dTB-BMPs); and 4) freshly isolated nondecellularized natural TBs (nTBs). Replicate samples were implanted into the mandibles of host Yucatan mini-pigs and grown for 3 or 6 mo. Harvested mandibles with implanted TB constructs were fixed in formalin, decalcified, embedded in paraffin, sectioned, and analyzed via histological methods. Micro-computed tomography (CT) analysis was performed on harvested 6-mo samples prior to decalcification. All harvested constructs exhibited a high degree of cellularity. Significant production of organized dentin and enamel-like tissues was observed in dTB-recell and nTB implants, but not in dTB or dTB-BMP implants. Micro-CT analyses of 6-mo implants showed the formation of organized, bioengineered teeth of comparable size to natural teeth. To our knowledge, these results are the first to describe the potential use of dTBs for functional whole tooth regeneration.

  1. Transcriptome analysis of chestnut (Castanea sativa) tree buds suggests a putative role for epigenetic control of bud dormancy.

    Science.gov (United States)

    Santamaría, María Estrella; Rodríguez, Roberto; Cañal, María Jesús; Toorop, Peter E

    2011-09-01

    Recent papers indicated that epigenetic control is involved in transitions in bud dormancy, purportedly controlling gene expression. The present study aimed to identify genes that are differentially expressed in dormant and non-dormant Castanea sativa buds. Two suppression subtractive hybridization cDNA libraries were constructed to characterize the transcriptomes of dormant apical buds of C. sativa, and buds in which dormancy was released. A total of 512 expressed sequence tags (ESTs) were generated in a forward and reverse subtractive hybridization experiment. Classification of these ESTs into functional groups demonstrated that dormant buds were predominantly characterized by genes associated with stress response, while non-dormant buds were characterized by genes associated with energy, protein synthesis and cellular components for development and growth. ESTs for a few genes involved in different forms of epigenetic modification were found in both libraries, suggesting a role for epigenetic control in bud dormancy different from that in growth. Genes encoding histone mono-ubiquitinase HUB2 and histone acetyltransferase GCN5L were associated with dormancy, while a gene encoding histone H3 kinase AUR3 was associated with growth. Real-time RT-PCR with a selection of genes involved in epigenetic modification and stress tolerance confirmed the expression of the majority of investigated genes in various stages of bud development, revealing a cyclical expression pattern concurring with the growth seasons for most genes. However, senescing leaves also showed an increased expression of several of the genes associated with dormancy, implying pleiotropy. Furthermore, a comparison between these subtraction cDNA libraries and the poplar bud dormancy transcriptome and arabidopsis transcriptomes for seed dormancy and non-dormancy indicated a common basis for dormancy in all three systems. Bud dormancy and non-dormancy in C. sativa were characterized by distinct sets of genes

  2. Taste bud homeostasis in health, disease, and aging.

    Science.gov (United States)

    Feng, Pu; Huang, Liquan; Wang, Hong

    2014-01-01

    The mammalian taste bud is an onion-shaped epithelial structure with 50-100 tightly packed cells, including taste receptor cells, supporting cells, and basal cells. Taste receptor cells detect nutrients and toxins in the oral cavity and transmit the sensory information to gustatory nerve endings in the buds. Supporting cells may play a role in the clearance of excess neurotransmitters after their release from taste receptor cells. Basal cells are precursor cells that differentiate into mature taste cells. Similar to other epithelial cells, taste cells turn over continuously, with an average life span of about 8-12 days. To maintain structural homeostasis in taste buds, new cells are generated to replace dying cells. Several recent studies using genetic lineage tracing methods have identified populations of progenitor/stem cells for taste buds, although contributions of these progenitor/stem cell populations to taste bud homeostasis have yet to be fully determined. Some regulatory factors of taste cell differentiation and degeneration have been identified, but our understanding of these aspects of taste bud homoeostasis remains limited. Many patients with various diseases develop taste disorders, including taste loss and taste distortion. Decline in taste function also occurs during aging. Recent studies suggest that disruption or alteration of taste bud homeostasis may contribute to taste dysfunction associated with disease and aging.

  3. Molecular and Pathogenetic Aspects of Tumor Budding in Colorectal Cancer

    Science.gov (United States)

    Dawson, Heather; Lugli, Alessandro

    2015-01-01

    In recent years, tumor budding in colorectal cancer has gained much attention as an indicator of lymph node metastasis, distant metastatic disease, local recurrence, worse overall and disease-free survival, and as an independent prognostic factor. Tumor buds, defined as the presence of single tumor cells or small clusters of up to five tumor cells at the peritumoral invasive front (peritumoral buds) or within the main tumor body (intratumoral buds), are thought to represent the morphological correlate of cancer cells having undergone epithelial–mesenchymal transition (EMT), an important mechanism for the progression of epithelial cancers. In contrast to their undisputed prognostic power and potential to influence clinical management, our current understanding of the biological background of tumor buds is less established. Most studies examining tumor buds have attempted to recapitulate findings of mechanistic EMT studies using immunohistochemical markers. The aim of this review is to provide a comprehensive summary of studies examining protein expression profiles of tumor buds and to illustrate the molecular pathways and crosstalk involved in their formation and maintenance. PMID:25806371

  4. Molecular and pathogenetic aspects of tumor budding in colorectal cancer

    Directory of Open Access Journals (Sweden)

    Heather eDawson

    2015-03-01

    Full Text Available In recent years, tumor budding in colorectal cancer has gained much attention as an indicator of lymph node metastasis, distant metastatic disease, local recurrence, worse overall and disease-free survival and as an independent prognostic factor. Tumor buds, defined as the presence of single tumor cells or small clusters of up to 5 tumor cells at the peritumoral invasive front (peritumoral buds or within the main tumor body (intratumoral buds, are thought to represent the morphological correlate of cancer cells having undergone epithelial-mesenchymal transition (EMT, an important mechanism for the progression of epithelial cancers. In contrast to their undisputed prognostic power and potential to influence clinical management, our current understanding of the biological background of tumor buds is less established. Most studies examining tumor buds have attempted to recapitulate findings of mechanistic EMT studies using immunohistochemical markers. The aim of this review is to provide a comprehensive summary of studies examining protein expression profiles of tumor buds and to illustrate the molecular pathways and crosstalk involved in their formation and maintenance.

  5. Budding & Grafting Citrus & Avocados in the Home Garden

    OpenAIRE

    Elam, Pam

    1997-01-01

    It is often tempting, after eating a particularly good orange or avocado, to plant the seed and grow your own tree full of these delicious fruit. The best way to produce good-quality fruit is to grow seedlings from them and then attach, by budding or grafting, material from trees that are known to be good producers. Budding and grafting can also be used to change or add varieties to mature citrus or avocado trees, a process known as top working. This publication is a brief introduction to bud...

  6. Factors Intrinsic to the Axillary Bud Determine Topophysic Effects on Bud and Shoot Growth and Flower Development in Rosa hybrida.

    Science.gov (United States)

    Bredmose; Hansen; Nielsen

    1999-09-01

    Topophysis (the influence of the position of axillary buds along the shoot on bud and shoot growth, fresh biomass accumulation, and flower development) were studied in Rosa hybrida L. Single-node cuttings with five-leaflet leaves were excised from nine stem positions and grown as single-stemmed rose plants. Plants were grown at 20-h photoperiods, 22 degrees C average air temperature, and an average photosynthetic photon flux density of 228 µmol m-2 s-1. Generally, onset of axillary bud growth and initial shoot growth were promoted, and flower height and bloom quality were reduced in plants from apical bud positions. Stem diameter, stem growth rate, and biomass buildup were greatest from medial bud positions; the plastochron was greater, and stems and internodes were longer, from basal bud positions. Flower diameter and cut flower vase life were not significantly influenced by topophysis. The results demonstrate that topophysis can determine the potential for plant development and flowering in R. hybrida and that this knowledge may be used to regulate plant growth. Results also indicate intrinsic mechanisms determining axillary bud growth, but the physiology of topophysis is presently unclear. A probable role of cytokinins in the topophysic prevention of growth observed is discussed.

  7. A novel role for the GTPase-activating protein Bud2 in the spindle position checkpoint.

    Directory of Open Access Journals (Sweden)

    Scott A Nelson

    Full Text Available The spindle position checkpoint (SPC ensures correct mitotic spindle position before allowing mitotic exit in the budding yeast Saccharomyces cerevisiae. In a candidate screen for checkpoint genes, we identified bud2Δ as deficient for the SPC. Bud2 is a GTPase activating protein (GAP, and the only known substrate of Bud2 was Rsr1/Bud1, a Ras-like GTPase and a central component of the bud-site-selection pathway. Mutants lacking Rsr1/Bud1 had no checkpoint defect, as did strains lacking and overexpressing Bud5, a guanine-nucleotide exchange factor (GEF for Rsr1/Bud1. Thus, the checkpoint function of Bud2 is distinct from its role in bud site selection. The catalytic activity of the Bud2 GAP domain was required for the checkpoint, based on the failure of the known catalytic point mutant Bud2(R682A to function in the checkpoint. Based on assays of heterozygous diploids, bud2(R682A, was dominant for loss of checkpoint but recessive for bud-site-selection failure, further indicating a separation of function. Tem1 is a Ras-like protein and is the critical regulator of mitotic exit, sitting atop the mitotic exit network (MEN. Tem1 is a likely target for Bud2, supported by genetic analyses that exclude other Ras-like proteins.

  8. Apoptosis at inflection point in liquid culture of budding yeasts.

    Directory of Open Access Journals (Sweden)

    Toshiyuki Hagiwara

    Full Text Available Budding yeasts are highly suitable for aging studies, because the number of bud scars (stage proportionally correlates with age. Its maximum stages are known to reach at 20-30 stages on an isolated agar medium. However, their stage dynamics in a liquid culture is virtually unknown. We investigate the population dynamics by counting scars in each cell. Here one cell division produces one new cell and one bud scar. This simple rule leads to a conservation law: "The total number of bud scars is equal to the total number of cells." We find a large discrepancy: extremely fewer cells with over 5 scars than expected. Almost all cells with 6 or more scars disappear within a short period of time in the late log phase (corresponds to the inflection point. This discrepancy is confirmed directly by the microscopic observations of broken cells. This finding implies apoptosis in older cells (6 scars or more.

  9. Ubiquitin is part of the retrovirus budding machinery

    Science.gov (United States)

    Patnaik, Akash; Chau, Vincent; Wills, John W.

    2000-11-01

    Retroviruses contain relatively large amounts of ubiquitin, but the significance of this finding has been unknown. Here, we show that drugs that are known to reduce the level of free ubiquitin in the cell dramatically reduced the release of Rous sarcoma virus, an avian retrovirus. This effect was suppressed by overexpressing ubiquitin and also by directly fusing ubiquitin to the C terminus of Gag, the viral protein that directs budding and particle release. The block to budding was found to be at the plasma membrane, and electron microscopy revealed that the reduced level of ubiquitin results in a failure of mature virus particles to separate from each other and from the plasma membrane during budding. These data indicate that ubiquitin is actually part of the budding machinery.

  10. Tolerance of budding yeast Saccharomyces cerevisiae to ultra high pressure

    Science.gov (United States)

    Shibata, M.; Torigoe, M.; Matsumoto, Y.; Yamamoto, M.; Takizawa, N.; Hada, Y.; Mori, Y.; Takarabe, K.; Ono, F.

    2014-05-01

    Our studies on the tolerance of plants and animals against very high pressure of several GPa have been extended to a smaller sized fungus, the budding yeast Saccharomyces cerevisiae. Several pieces of budding yeast (dry yeast) were sealed in a small teflon capsule with a liquid pressure medium fluorinate, and exposed to 7.5 GPa by using a cubic anvil press. The pressure was kept constant for various duration of time from 2 to 24 h. After the pressure was released, the specimens were brought out from the teflon capsule, and they were cultivated on a potato dextrose agar. It was found that the budding yeast exposed to 7.5 GPa for up to 6 h showed multiplication. However, those exposed to 7.5 GPa for longer than 12 h were found dead. The high pressure tolerance of budding yeast is a little weaker than that of tardigrades.

  11. Tumor Budding in Upper Gastrointestinal Carcinomas

    Science.gov (United States)

    Koelzer, Viktor H.; Langer, Rupert; Zlobec, Inti; Lugli, Alessandro

    2014-01-01

    The basis of personalized medicine in oncology is the prediction of an individual’s risk of relapse and death from disease. The presence of tumor budding (TB) at the tumor–host interface of gastrointestinal cancers has been recognized as a hallmark of unfavorable disease biology. TB is defined as the presence of dedifferentiated cells or small clusters of up to five cells at the tumor invasive front and can be observed in aggressive carcinomas of the esophagus, stomach, pancreas, ampulla, colon, and rectum. Presence of TB reproducibly correlates with advanced tumor stage, frequent lymphovascular invasion, nodal, and distant metastasis. The UICC has officially recognized TB as additional independent prognostic factor in cancers of the colon and rectum. Recent studies have also characterized TB as a promising prognostic indicator for clinical management of esophageal squamous cell carcinoma, adenocarcinoma of the gastro-esophageal junction, and gastric adenocarcinoma. However, several important issues have to be addressed for application in daily diagnostic practice: (1) validation of prognostic scoring systems for TB in large, multi-center studies, (2) consensus on the optimal assessment method, and (3) inter-observer reproducibility. This review provides a comprehensive analysis of TB in cancers of the upper gastrointestinal tract including critical appraisal of perspectives for further study. PMID:25177546

  12. Taste Bud Homeostasis in Health, Disease, and Aging

    OpenAIRE

    Feng, Pu; Huang, Liquan; Wang, Hong

    2013-01-01

    The mammalian taste bud is an onion-shaped epithelial structure with 50–100 tightly packed cells, including taste receptor cells, supporting cells, and basal cells. Taste receptor cells detect nutrients and toxins in the oral cavity and transmit the sensory information to gustatory nerve endings in the buds. Supporting cells may play a role in the clearance of excess neurotransmitters after their release from taste receptor cells. Basal cells are precursor cells that differentiate into mature...

  13. Longleaf pine bud development: influence of seedling nutrition

    Science.gov (United States)

    J. P. Barnett; D. P. Jackson; R. K. Dumroese

    2010-01-01

    A subset of seedlings from a larger study (Jackson and others 2006, 2007) were selected and evaluated for two growing seasons to relate bud development, and root-collar diameter (RCD), and height growth with three nursery fertilization rates. We chose seedlings in the 0.5 (lowest), 2.0 (mid-range), and 4.0 (highest) mg of nitrogen per seedling treatments. Buds moved...

  14. Real Life Science with Dandelions and Project BudBurst.

    Science.gov (United States)

    Johnson, Katherine A

    2016-03-01

    Project BudBurst is a national citizen-science project that tracks bloom times and other phenological data for plants across the country. Data from Project BudBurst are being used to measure the effects of climate change. Students can participate in this project by watching any of the plants on the list, including the common dandelion, which makes the program easy and accessible to everyone. Journal of Microbiology & Biology Education.

  15. Real Life Science with Dandelions and Project BudBurst

    Directory of Open Access Journals (Sweden)

    Katherine A. Johnson

    2015-12-01

    Full Text Available Project BudBurst is a national citizen-science project that tracks bloom times and other phenological data for plants across the country. Data from Project BudBurst are being used to measure the effects of climate change. Students can participate in this project by watching any of the plants on the list, including the common dandelion, which makes the program easy and accessible to everyone.

  16. Tumor Budding: The Name is EMT. Partial EMT.

    Science.gov (United States)

    Grigore, Alexandru Dan; Jolly, Mohit Kumar; Jia, Dongya; Farach-Carson, Mary C; Levine, Herbert

    2016-04-29

    Tumor budding is a histological phenomenon encountered in various cancers, whereby individual malignant cells and/or small clusters of malignant cells are seen in the tumor stroma. Postulated to be mirror epithelial-mesenchymal transition, tumor budding has been associated with poor cancer outcomes. However, the vast heterogeneity in its exact definition, methodology of assessment, and patient stratification need to be resolved before it can be routinely used as a standardized prognostic feature. Here, we discuss the heterogeneity in defining and assessing tumor budding, its clinical significance across multiple cancer types, and its prospective implementation in clinical practice. Next, we review the emerging evidence about partial, rather than complete, epithelial-mesenchymal phenotype at the tumor bud level, and its connection with tumor proliferation, quiescence, and stemness. Finally, based on recent literature, indicating a co-expression of epithelial and mesenchymal markers in many tumor buds, we posit tumor budding to be a manifestation of this hybrid epithelial/mesenchymal phenotype displaying collective cell migration.

  17. Tumor Budding: The Name is EMT. Partial EMT.

    Directory of Open Access Journals (Sweden)

    Alexandru Dan Grigore

    2016-04-01

    Full Text Available Tumor budding is a histological phenomenon encountered in various cancers, whereby individual malignant cells and/or small clusters of malignant cells are seen in the tumor stroma. Postulated to be mirror epithelial-mesenchymal transition, tumor budding has been associated with poor cancer outcomes. However, the vast heterogeneity in its exact definition, methodology of assessment, and patient stratification need to be resolved before it can be routinely used as a standardized prognostic feature. Here, we discuss the heterogeneity in defining and assessing tumor budding, its clinical significance across multiple cancer types, and its prospective implementation in clinical practice. Next, we review the emerging evidence about partial, rather than complete, epithelial-mesenchymal phenotype at the tumor bud level, and its connection with tumor proliferation, quiescence, and stemness. Finally, based on recent literature, indicating a co-expression of epithelial and mesenchymal markers in many tumor buds, we posit tumor budding to be a manifestation of this hybrid epithelial/mesenchymal phenotype displaying collective cell migration.

  18. Tumor budding in colorectal carcinoma: time to take notice.

    Science.gov (United States)

    Mitrovic, Bojana; Schaeffer, David F; Riddell, Robert H; Kirsch, Richard

    2012-10-01

    Tumor 'budding', loosely defined by the presence of individual cells and small clusters of tumor cells at the invasive front of carcinomas, has received much recent attention, particularly in the setting of colorectal carcinoma. It has been postulated to represent an epithelial-mesenchymal transition. Tumor budding is a well-established independent adverse prognostic factor in colorectal carcinoma that may allow for stratification of patients into risk categories more meaningful than those defined by TNM staging, and also potentially guide treatment decisions, especially in T1 and T3 N0 (Stage II, Dukes' B) colorectal carcinoma. Unfortunately, its universal acceptance as a reportable factor has been held back by a lack of definitional uniformity with respect to both qualitative and quantitative aspects of tumor budding. The purpose of this review is fourfold: (1) to describe the morphology of tumor budding and its relationship to other potentially important features of the invasive front; (2) to summarize current knowledge regarding the prognostic significance and potential clinical implications of this histomorphological feature; (3) to highlight the challenges posed by a lack of data to allow standardization with respect to the qualitative and quantitative criteria used to define budding; and (4) to present a practical approach to the assessment of tumor budding in everyday practice.

  19. Tumor budding correlates with poor prognosis and epithelial-mesenchymal transition in tongue squamous cell carcinoma.

    Science.gov (United States)

    Wang, Cheng; Huang, Hongzhang; Huang, Zhiquan; Wang, Anxun; Chen, Xiaohua; Huang, Lei; Zhou, Xiaofeng; Liu, Xiqiang

    2011-08-01

    Tumor budding is a readily detectable histopathological feature and has been recognized as an adverse prognostic factor in several human cancers. However, the prognostic value of tumor budding in tongue squamous cell carcinoma (TSCC) has not been reported. The purpose of this study was to assess the correlation of tumor budding with the clinicopathologic features, and the known molecular biomarkers (E-cadherin and Vimentin), as well as to evaluate its prognostic significance for TSCC. Archival clinical samples of 230 patients with TSCC were examined for tumor budding. Immunohistochemistry analyses were performed to examine the expression of E-cadherin and Vimentin. Statistical analyses were carried out to assess the correlation of tumor budding with clinicopathologic parameters and patient survival. The potential association between tumor budding and alterations of E-cadherin and Vimentin expression was also assessed. Of the 230 TSCC cases examined, tumor budding was observed in 165 cases (71.7%), with a mean tumor bud count of 7.5 (range from 1 to 48 buds). High-intensity budding (≥5 tumor buds) was observed in 111 cases (48.3%). Statistical analysis revealed that tumor budding was associated with tumor size (P tumor budding and the deregulation of E-cadherin (P Tumor budding, which associates with epithelial-mesenchymal transition, is a frequent event and appears to be an independent prognostic factor in TSCC. © 2011 John Wiley & Sons A/S.

  20. Electron tomography reveals the steps in filovirus budding.

    Directory of Open Access Journals (Sweden)

    Sonja Welsch

    2010-04-01

    Full Text Available The filoviruses, Marburg and Ebola, are non-segmented negative-strand RNA viruses causing severe hemorrhagic fever with high mortality rates in humans and nonhuman primates. The sequence of events that leads to release of filovirus particles from cells is poorly understood. Two contrasting mechanisms have been proposed, one proceeding via a "submarine-like" budding with the helical nucleocapsid emerging parallel to the plasma membrane, and the other via perpendicular "rocket-like" protrusion. Here we have infected cells with Marburg virus under BSL-4 containment conditions, and reconstructed the sequence of steps in the budding process in three dimensions using electron tomography of plastic-embedded cells. We find that highly infectious filamentous particles are released at early stages in infection. Budding proceeds via lateral association of intracellular nucleocapsid along its whole length with the plasma membrane, followed by rapid envelopment initiated at one end of the nucleocapsid, leading to a protruding intermediate. Scission results in local membrane instability at the rear of the virus. After prolonged infection, increased vesiculation of the plasma membrane correlates with changes in shape and infectivity of released viruses. Our observations demonstrate a cellular determinant of virus shape. They reconcile the contrasting models of filovirus budding and allow us to describe the sequence of events taking place during budding and release of Marburg virus. We propose that this represents a general sequence of events also followed by other filamentous and rod-shaped viruses.

  1. Electron tomography reveals the steps in filovirus budding.

    Science.gov (United States)

    Welsch, Sonja; Kolesnikova, Larissa; Krähling, Verena; Riches, James D; Becker, Stephan; Briggs, John A G

    2010-04-29

    The filoviruses, Marburg and Ebola, are non-segmented negative-strand RNA viruses causing severe hemorrhagic fever with high mortality rates in humans and nonhuman primates. The sequence of events that leads to release of filovirus particles from cells is poorly understood. Two contrasting mechanisms have been proposed, one proceeding via a "submarine-like" budding with the helical nucleocapsid emerging parallel to the plasma membrane, and the other via perpendicular "rocket-like" protrusion. Here we have infected cells with Marburg virus under BSL-4 containment conditions, and reconstructed the sequence of steps in the budding process in three dimensions using electron tomography of plastic-embedded cells. We find that highly infectious filamentous particles are released at early stages in infection. Budding proceeds via lateral association of intracellular nucleocapsid along its whole length with the plasma membrane, followed by rapid envelopment initiated at one end of the nucleocapsid, leading to a protruding intermediate. Scission results in local membrane instability at the rear of the virus. After prolonged infection, increased vesiculation of the plasma membrane correlates with changes in shape and infectivity of released viruses. Our observations demonstrate a cellular determinant of virus shape. They reconcile the contrasting models of filovirus budding and allow us to describe the sequence of events taking place during budding and release of Marburg virus. We propose that this represents a general sequence of events also followed by other filamentous and rod-shaped viruses.

  2. Budżet zadaniowy w oświacie

    OpenAIRE

    Kopańska, Agnieszka

    2012-01-01

    Budżet to podstawowe narzędzie zarządzania finansami publicznymi. Jego forma – a więc to jak poszczególne wydatki i dochody są w nim zapisywane stanowi jeden z najważniejszych czynników wpływających na efektywność publicznych podmiotów. W ostatnich dziesięcioleciach, coraz częściej postuluje się odejście od klasycznej formy budżetu i poszukuje się takich które tę efektywność będą wyraźnie poprawiać. W niniejszym opracowaniu analizie poddane są możliwości i ograniczenia zastosowania budże...

  3. Adventitious bud regeneration from the stigma of Sinapis alba L.

    Directory of Open Access Journals (Sweden)

    Elżbieta Zenkteler

    2012-12-01

    Full Text Available Stigmas isolated from flower buds of 'Nakielska' variety of Sinapis alba were used to develop a micropropagation method suitable for breeding of new cultivars. The origin of adventitious bud regeneration was studied on MS medium, under stimulation by bezylaminopurine (BAP in combination with 2,4-D - dichlorophenoxyacetic acid (2,4-D. Histological analysis showed the structure of Sinapis stigma (composed from four types of tissue: papillae, transmitting tissue, parenchyma and vascular bundles and revealed that numerous meristematic centers developed from parenchyma cells in close vicinity of vascular bundles. Buds very quickly appeared on the surface of initial explants and later formed multiplantlets that were easily rooted in the soil.

  4. EARLY BUD-BREAK1 (EBB1) defines a conserved mechanism for control of bud-break in woody perennials.

    Science.gov (United States)

    Busov, Victor; Carneros, Elena; Yakovlev, Igor

    2016-01-01

    Bud-break is an environmentally and economically important trait in trees, shrubs and vines from temperate latitudes. Poor synchronization of bud-break timing with local climates can lead to frost injuries, susceptibility to pests and pathogens and poor crop yields in fruit trees and vines. The rapid climate changes outpace the adaptive capacities of plants to respond through natural selection. This is particularly true for trees which have long generation cycle and thus the adaptive changes are significantly delayed. Therefore, to devise appropriate breeding and conservation strategies, it is imperative to understand the molecular underpinnings that govern dormancy mechanisms. We have recently identified and characterized the poplar EARLY BUD-BREAK 1 (EBB1) gene. EBB1 is a positive regulator of bud-break and encodes a transcription factor from the AP2/ERF family. Here, using comparative and functional genomics approaches we show that EBB1 function in regulation of bud-break is likely conserved across wide range of woody perennial species with importance to forestry and agriculture.

  5. Dormancy in Peach (Prunus persica L.) Flower Buds 1

    Science.gov (United States)

    Luna, Virginia; Lorenzo, Eugenia; Reinoso, Herminda; Tordable, Maria C.; Abdala, Guillermina; Pharis, Richard P.; Bottini, Ruben

    1990-01-01

    Flower buds of peach (Prunus persica L.) trees, cv Novedad de Cordoba (Argentina), were collected near the end of the dormant period and immediately before anthesis. After removal of scale leaves, morphological observations of representative buds, made on transverse and longitudinal microtome sections, showed that all verticils making up the flower are present in an undifferentiated form during the dormant period (June). Flower buds collected at the end of dormant period (August) showed additional growth and differentiation, at which time formation of two ovules was beginning in the unicarpelar gynoecium. Dehiscence of anthers had not yet occurred 10 days before full bloom, and the ovules were still developing. Free endogenous gibberellin (GA)-like substances were quantified by bioassay (Tan-ginbozu dwarf rice microdrop) after SiO2 partition column chromatography, reversed phase C18-high performance liquid chromatography, and finally Nucleosil [N(CH3)2]high performance liquid chromatography. Bioactive fractions were then subjected to capillary gas chromatography-mass spectrometry-selected ion monitoring (GC-MS-SIM). Gibberellins A1, A3, and A8 were tentatively identified in peach flower buds using GC-SIM and Kovat's retention indices, and relative amounts approximated by GC-SIM (2:8:6 for GA1, GA3, and GA8, respectively). The highest concentration (330 nanograms per gram dry weight) of free GA1/GA3 was found in dormant buds (June) and diminished thereafter. The concentration free of GA1/GA3 did not increase immediately prior to bud break. However, high GA1/GA3 concentrations occurred during stages where rate of growth and cellular differentiation of (mainly fertile) verticils can be influenced. Images Figure 1 PMID:16667435

  6. Histopathologic features of the tumor budding in adenocarcinoma of the lung: tumor budding as an index to predict the potential aggressiveness.

    Science.gov (United States)

    Yamaguchi, Yoko; Ishii, Genichiro; Kojima, Motohiro; Yoh, Kiyotaka; Otsuka, Hajime; Otaki, Yoichi; Aokage, Keiju; Yanagi, Shingo; Nagai, Kanji; Nishiwaki, Yutaka; Ochiai, Atsushi

    2010-09-01

    The term tumor budding has been applied to single cells or small clusters of up to four cells within the stromal tissue at the invasive margin of colorectal cancers. This morphologic feature is increasingly being recognized as an adverse prognostic factor. The purpose of this study was to evaluate the clinicopathologic significance of tumor budding in adenocarcinomas of the lung. We investigated the relationship between tumor budding and clinicopathologic parameters of adenocarcinomas of the lung and the prognostic significance of tumor budding by reviewing the cases of 201 consecutive patients who had undergone complete resection of adenocarcinoma of the lung measuring 30 mm or less in diameter. We examined immunohistochemical profile of budding cells (BCs) by immunohistochemical staining with 14 antibodies. Tumor budding was observed in 78 (43.1%) of the 181 cases with invasive adenocarcinoma. The presence of tumor budding was significantly associated with lymph node metastasis (p = 0.005), pathologic stage (p tumor budding and the predominant histologic subtype revealed that the predominant papillary subtype was significantly associated with the presence of tumor budding (p = 0.0023), whereas the predominant bronchioloalveolar carcinoma subtype was significantly associated with the absence of tumor budding (p budding and the group without budding was 67.5% and 88.3%, respectively, and difference was significant (p = 0.0057). Compared with cancer cells forming nests, BCs displayed reduced expression of cellular adhesion molecule, E-cadherin, and beta-catenin (p tumor budding was significant independent prognostic factor of the small-sized adenocarcinoma of the lung. Our data showed that tumor budding in adenocarcinoma of the lung is a distinct morphologic feature that has biologic and prognostic significance.

  7. Effect of alternating day and night temperature on short day-induced bud set and subsequent bud burst in long days in Norway spruce

    Directory of Open Access Journals (Sweden)

    Jorunn Elisabeth Olsen

    2014-12-01

    Full Text Available Young seedlings of the conifer Norway spruce exhibit short day (SD-induced cessation of apical growth and bud set. Although different, constant temperatures under SD are known to modulate timing of bud set and depth of dormancy with development of deeper dormancy under higher compared to lower temperature, systematic studies of effects of alternating day (DT and night temperatures (NT are limited. To shed light on this, seedlings of different provenances of Norway spruce were exposed to a wide range of DT-NT combinations during bud development, followed by transfer to forcing conditions of long days (LD and 18°C, directly or after different periods of chilling. Although no specific effect of alternating DT/NT was found, the results demonstrate that the effects of DT under SD on bud set and subsequent bud break are significantly modified by NT in a complex way. The effects on bud break persisted after chilling. Since time to bud set correlated with the daily mean temperature under SD at DTs of 18 and 21°C, but not a DT of 15°C, time to bud set apparently also depend on the specific DT, implying that the effect of NT depends on the actual DT. Although higher temperature under SD generally results in later bud break after transfer to forcing conditions, the fastest bud flush was observed at intermediate NTs. This might be due to a bud break-hastening chilling effect of intermediate compared to higher temperatures, and delayed bud development to a stage where bud burst can occur, under lower temperatures. Also, time to bud burst in un-chilled seedlings decreased with increasing SD-duration, suggesting that bud development must reach a certain stage before the processes leading to bud burst are initiated. The present results also indicate that low temperature during bud development had a larger effect on the most southern compared to the most northern provenance studied. Decreasing time to bud burst was observed with increasing northern latitude

  8. The influence of gravity on bud development in apple trees and in poplars

    Directory of Open Access Journals (Sweden)

    B. Borkowska

    2015-06-01

    Full Text Available The lower shoots in horizontally placed apple trees exhibited much weaker development as well in light as darkness. Removing a narrow strip of bark along both sides of a horizontally placed apple tree improved markedly the growth of the lower buds. In poplars the same effect was received by surrounding the lower buds with semicircular incisions. The inhibition of the lower buds was also released by removing the apical bud and the upper ones. The presented results and those published earlier show that the mechanism responsible for inhibition of the lower buds acts in two steps: 1 gravity influences directly the system composed of a bud and the adjacent tissue of the stem. 2 the lower buds partly inhibited in the step "1" are further inhibited by a correlative mechanism which supresses all weaker buds. The second "step" reaction takes place also after a tree have been reverted back to the normal vertical position.

  9. Extensive transcriptome changes during natural onset and release of vegetative bud dormancy in Populus

    Science.gov (United States)

    To survive winter conditions, axillary buds of poplar transition from paradormancy to endodormancy. Following sufficient chilling, endodormant axillary buds will transition from endodormancy to ecodormancy. We utilized the near whole genome NimbleGen poplar microarrays to follow transcriptome diff...

  10. β-Catenin signaling regulates temporally discrete phases of anterior taste bud development

    OpenAIRE

    Thirumangalathu, Shoba; Barlow, Linda A.

    2015-01-01

    The sense of taste is mediated by multicellular taste buds located within taste papillae on the tongue. In mice, individual taste buds reside in fungiform papillae, which develop at mid-gestation as epithelial placodes in the anterior tongue. Taste placodes comprise taste bud precursor cells, which express the secreted factor sonic hedgehog (Shh) and give rise to taste bud cells that differentiate around birth. We showed previously that epithelial activation of β-catenin is the primary induct...

  11. The influence of gravity on bud development in apple trees and in poplars

    OpenAIRE

    B. Borkowska; L. S. Jankiewicz

    2015-01-01

    The lower shoots in horizontally placed apple trees exhibited much weaker development as well in light as darkness. Removing a narrow strip of bark along both sides of a horizontally placed apple tree improved markedly the growth of the lower buds. In poplars the same effect was received by surrounding the lower buds with semicircular incisions. The inhibition of the lower buds was also released by removing the apical bud and the upper ones. The presented results and those published earlier s...

  12. In vitro PROLIFERATION ABILITY OF AXILLARY BUDS IN Musa spp

    African Journals Online (AJOL)

    AISA

    E-mail : youmbi_emmanuel@yahoo.fr. 2University of Yaounde I, Faculty of Sciences, Laboratory of Biotechnology and Environment, Unit of Physiology and Plant Breeding. B.P. 812 Yaounde Cameroon. ABSTRACT. Tissue culture method has always considered the apical bud as the initial explant for micropropagation of.

  13. presence of axillary bud and application of plant growth hormones ...

    African Journals Online (AJOL)

    D. alata)- were grown in pots in the greenhouse. Half the cuttings bore axillary buds and half had none. The cuttings were sprayed with a factorial combination of indole acetic acid (IAA), benzyladenine (BA) and giberellic acid (GA3). Cuttings ...

  14. thidiazuron improves adventitious bud and shoot regeneration in ...

    African Journals Online (AJOL)

    Prof. Adipala Ekwamu

    Induction of adventitious buds and shoots from intact leaves and stem internode segments of two recalcitrant. Ugandan sweetpotato (Ipomoea batatas L.) cultivars was investigated in vitro on Murashige and Skoog (MS) medium, supplemented with 3 different levels (0.5, 2.0 and 4.0 µM) of Thidiazuron (TDZ). Shoots were.

  15. Regulation of homologous recombination at telomeres in budding yeast

    DEFF Research Database (Denmark)

    Eckert-Boulet, Nadine; Lisby, Michael

    2010-01-01

    Homologous recombination is suppressed at normal length telomere sequences. In contrast, telomere recombination is allowed when telomeres erode in the absence of telomerase activity or as a consequence of nucleolytic degradation or incomplete replication. Here, we review the mechanisms...... that contribute to regulating mitotic homologous recombination at telomeres and the role of these mechanisms in signalling short telomeres in the budding yeast Saccharomyces cerevisiae....

  16. Dynamic assessment of Capparis spinosa buds on survival of ...

    African Journals Online (AJOL)

    Dynamic assessment of Capparis spinosa buds on survival of periodontal ligament cells using a real‑time cell analysis method. ... Results: Dulbecco's Modified Eagle Medium (control) and C. spinosa groups had significantly higher cell index values compared with the HBSS and light milk (P < 0.05). Although, C. spinosa ...

  17. In vitro multiple shoot bud induction and regeneration from plumule ...

    African Journals Online (AJOL)

    The response of eleven Indian cultivars of pigeon pea for in vitro multiple shoot bud induction and regeneration from plumule junction explants under variable concentration of 6-benzyl amino purine (BAP), kinetin and thiadiazuron (TDZ) was assessed in the present study. The cultivar IPA-3088 showed best response with a ...

  18. Effects of population source and node position on rhizome bud ...

    African Journals Online (AJOL)

    The effect of population source (agro-ecozone) and node position on bud distribution in mature (determinate) rhizome of first- (from natural vegetati on) and second- (from screenhouse-grown plants) generation speargrass (Imperata cylindrica (L.) Raeuschel) from the derived savanna (DS), southern Guinea savanna (SGS) ...

  19. Extending the dormant bud cryopreservation method to new tree species

    Science.gov (United States)

    In cryopreservation of germplasm, using dormant winter buds (DB) as source plant material is economically favorable over tissue culture options. Although the DB cryopreservation method has been known for many years, the approach is feasible only for cryopreserving a select number of temperate tree s...

  20. Tumor Budding in Breast Carcinoma: Relation to E-Cadherin, MMP-9 Expression, and Metastasis Risk

    Directory of Open Access Journals (Sweden)

    Ni Putu Sriwidyani

    2016-10-01

    Full Text Available Background: Tumor budding is a histopathologic entity refers to small cluster of cancer cells at the invasive edge of tumor. It was assumed that tumor budding is linked to epithelial-mesenchymal transition, an early event in metastasis. Objective: This study aimed to find out the correlation of tumor budding with E-cadherin and MMP-9 expression and risk of metastasis in breast carcinoma. Method: We investigated 35 cases breast carcinoma with metastasis and 35 cases without metastasis. The number of tumor budding was counted in cytokeratin-stained slides with 400x magnification (0.57 mm2. Result: Cut-off point by ROC analysis was 11 and the patient was categorized into low grade (0-10 buds and high grade (11 or more buds tumor budding. Inter-observer agreement was good with K value 0.914. Low level of E-cadherin was not significantly correlated with high grade tumor budding (p=0.660, meanwhile high level of MMP-9 was significantly correlated with high grade tumor budding (p=0.001. High grade tumor budding was a significant, independent risk factor of metastasis in breast carcinoma (OR=38.2, 95% CI 7.5-193.7, p<0.001. Conclusion: In conclusion, tumor budding grade is related to level of MMP-9 but has no correlation E-cadherin expression. High grade tumor budding is an independent risk factor of metastasis in breast carcinoma.

  1. Characterisation of flower bud opening in roses; a comparison of Madelon and Sonia roses

    NARCIS (Netherlands)

    Kuiper, D.; Reenen, van H.S.; Ribôt, S.A.

    1996-01-01

    Cut cv. Madelon rose buds do not open satisfactorily when kept under low light (LL) conditions, in contrast to cv. Sonia buds. Adding sucrose to the keeping solution or storage in high light (HL) helped Madelon buds to open, but probably two different mechanisms were involved. Satisfactory

  2. The occurrence of taste buds in adults of the terrestrial ceacilian ...

    African Journals Online (AJOL)

    Two generations of gustatory organs occur during amphibian ontogeny in frogs and salamanders (Anura and Caudata), and are classified as taste buds or taste discs. Taste buds are present in larval forms, whereas taste discs are typical for adults. The little research done on Gymnophiona suggests that only taste buds are ...

  3. Induction of ectopic taste buds by SHH reveals the competency and plasticity of adult lingual epithelium

    Science.gov (United States)

    Castillo, David; Seidel, Kerstin; Salcedo, Ernesto; Ahn, Christina; de Sauvage, Frederic J.; Klein, Ophir D.; Barlow, Linda A.

    2014-01-01

    Taste buds are assemblies of elongated epithelial cells, which are innervated by gustatory nerves that transmit taste information to the brain stem. Taste cells are continuously renewed throughout life via proliferation of epithelial progenitors, but the molecular regulation of this process remains unknown. During embryogenesis, sonic hedgehog (SHH) negatively regulates taste bud patterning, such that inhibition of SHH causes the formation of more and larger taste bud primordia, including in regions of the tongue normally devoid of taste buds. Here, using a Cre-lox system to drive constitutive expression of SHH, we identify the effects of SHH on the lingual epithelium of adult mice. We show that misexpression of SHH transforms lingual epithelial cell fate, such that daughter cells of lingual epithelial progenitors form cell type-replete, onion-shaped taste buds, rather than non-taste, pseudostratified epithelium. These SHH-induced ectopic taste buds are found in regions of the adult tongue previously thought incapable of generating taste organs. The ectopic buds are composed of all taste cell types, including support cells and detectors of sweet, bitter, umami, salt and sour, and recapitulate the molecular differentiation process of endogenous taste buds. In contrast to the well-established nerve dependence of endogenous taste buds, however, ectopic taste buds form independently of both gustatory and somatosensory innervation. As innervation is required for SHH expression by endogenous taste buds, our data suggest that SHH can replace the need for innervation to drive the entire program of taste bud differentiation. PMID:24993944

  4. Visible dormant buds as related to tree diameter and log position

    Science.gov (United States)

    H. Clay Smith

    1967-01-01

    Red oaks and yellow-poplars in a stand of second-growth cove hardwoods in West Virginia were studied to determine whether visible dormant buds are related to tree size or log position. No correlation was found between dormant buds and tree size, for either species; but yellow-poplars had a significantly greater number of buds on the upper log.

  5. Effects of bud loading levels and nitrogen doses on yield, physical ...

    African Journals Online (AJOL)

    The aim of this study was to investigate the effects of several bud loading levels in winter pruning and nitrogen doses on yield and physical and chemical properties of fresh vine-leaves of grape cultivar “Narince”. Vines trained with bilateral cordon system was pruned to yield 35000 to 53000 buds/ha (16 or 24 buds/vine) ...

  6. Effect of Cleopatra mandarin rootstock age on bud 'take' of Late ...

    African Journals Online (AJOL)

    Cleopatra mandarin rootstocks of ages 9 months, 10 months, 11 months and 12 months were budded with Late Valencia sweet orange variety using the chip budding technique in a randomised complete block design. There were 25 budded seedlings for each age group and replicated four times. Results obtained indicated ...

  7. Greater bud outgrowth of Bromus inermis than Pascopyrum smithii under multiple environmental conditions

    Science.gov (United States)

    Jacqueline P. Ott; Jack L. Butler; Yuping Rong; Lan. Xu

    2017-01-01

    Tiller recruitment of perennial grasses in mixed-grass prairie primarily occurs from belowground buds. Environmental conditions, such as temperature, soil moisture and grazing can affect bud outgrowth of both invasive and native perennial grasses. Differential bud outgrowth responses of native and invasive species to climate change and grazing could alter...

  8. Morphology and prognostic value of tumor budding in rectal cancer after neoadjuvant radiotherapy.

    Science.gov (United States)

    Du, Changzheng; Xue, Weicheng; Li, Jiyou; Cai, Yong; Gu, Jin

    2012-07-01

    Tumor budding is an acknowledged prognostic marker in colorectal cancer. This study was conducted to investigate the morphology and prognostic significance of budding in rectal cancer after neoadjuvant radiotherapy. Surgical specimens from 96 consecutive patients who underwent neoadjuvant radiotherapy and curative resection were retrieved to assess budding and other clinicopathologic factors. The morphology and prognostic significance of postirradiation tumor budding were closely associated with tumor regression grade. In the tumor regression grade 1 group, tumor budding presented as "false budding" and did not have a significant association with prognosis. In the tumor regression grade 2 and 3 groups, budding was observed surrounded by radiation-induced fibrosis and large populations of infiltrating inflammatory cells, and budding intensity was significantly associated with histologic differentiation, ypN stage, and lymphovascular invasion (P budding subgroup showed a significantly higher rate of 5-year disease-free survival than the high-grade budding subgroup (87.5% versus 55.6%, P tumor regression grade, and tumor budding were the major independent factors affecting long-term disease-free survival. In conclusion, postirradiation budding has distinct morphology and prognostic significance in rectal cancer after neoadjuvant radiotherapy. Copyright © 2012 Elsevier Inc. All rights reserved.

  9. Evaluation of twig pre-harvest temperature for effective cryopreservation of Vaccinium dormant buds

    Science.gov (United States)

    Cryopreservation of plant material by dormant buds is less expensive than using shoot tips; however currently, dormant buds are used only for preservation of selected temperate tree and shrub species. Using dormant buds could be an efficient strategy for long-term preservation of blueberry (Vacciniu...

  10. File list: NoD.Pan.50.AllAg.Pancreatic_bud [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  11. File list: ALL.Pan.10.AllAg.Pancreatic_bud [Chip-atlas[Archive

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  17. File list: ALL.Pan.50.AllAg.Pancreatic_bud [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  18. Influence of temperature on bud break, shoot growth, flower bud atrophy and winter production of glasshouse roses

    NARCIS (Netherlands)

    Berg, van den G.A.

    1987-01-01

    The influence of temperature in the range 15-22 °C on growth, production, quality and flower bud atrophy ('blindness') of the rose cultivars Sweet Promise and Varlon was studied. The roses were grown in Dutch glasshouse soil under natural light conditions and studied from October until May

  19. Membrane-elasticity model of Coatless vesicle budding induced by ESCRT complexes.

    Directory of Open Access Journals (Sweden)

    Bartosz Różycki

    Full Text Available The formation of vesicles is essential for many biological processes, in particular for the trafficking of membrane proteins within cells. The Endosomal Sorting Complex Required for Transport (ESCRT directs membrane budding away from the cytosol. Unlike other vesicle formation pathways, the ESCRT-mediated budding occurs without a protein coat. Here, we propose a minimal model of ESCRT-induced vesicle budding. Our model is based on recent experimental observations from direct fluorescence microscopy imaging that show ESCRT proteins colocalized only in the neck region of membrane buds. The model, cast in the framework of membrane elasticity theory, reproduces the experimentally observed vesicle morphologies with physically meaningful parameters. In this parameter range, the minimum energy configurations of the membrane are coatless buds with ESCRTs localized in the bud neck, consistent with experiment. The minimum energy configurations agree with those seen in the fluorescence images, with respect to both bud shapes and ESCRT protein localization. On the basis of our model, we identify distinct mechanistic pathways for the ESCRT-mediated budding process. The bud size is determined by membrane material parameters, explaining the narrow yet different bud size distributions in vitro and in vivo. Our membrane elasticity model thus sheds light on the energetics and possible mechanisms of ESCRT-induced membrane budding.

  20. Tumor budding is an independent adverse prognostic factor in pancreatic ductal adenocarcinoma.

    Science.gov (United States)

    O'Connor, Kate; Li-Chang, Hector H; Kalloger, Steven E; Peixoto, Renata D; Webber, Douglas L; Owen, David A; Driman, David K; Kirsch, Richard; Serra, Stefano; Scudamore, Charles H; Renouf, Daniel J; Schaeffer, David F

    2015-04-01

    Tumor budding is a well-established adverse prognostic factor in colorectal cancer. However, the significance and diagnostic reproducibility of budding in pancreatic carcinoma requires further study. We aimed to assess the prognostic significance of tumor budding in pancreatic ductal adenocarcinoma, determine its relationship with other clinicopathologic features, and assess interobserver variability in its diagnosis. Tumor budding was assessed in 192 archival cases of pancreatic ductal adenocarcinoma using hematoxylin and eosin (H&E) sections; tumor buds were defined as single cells or nonglandular clusters composed of budding was determined through assessment of all tumor-containing slides, and associations with clinicopathologic features and outcomes were analyzed. Six gastrointestinal pathologists participated in an interobserver variability study of 120 images of consecutive tumor slides stained with H&E and cytokeratin. Budding was present in 168 of 192 cases and was associated with decreased overall survival (P=0.001). On multivariable analysis, tumor budding was prognostically significantly independent of stage, grade, tumor size, nodal status, lymphovascular invasion, and perineural invasion. There was substantial agreement among pathologists in assessing the presence of tumor budding using both H&E (K=0.63) and cytokeratin (K=0.63) stains. The presence of tumor budding is an independent adverse prognostic factor in pancreatic ductal carcinoma. The assessment of budding with H&E is reliable and could be used to better risk stratify patients with pancreatic ductal adenocarcinoma.

  1. Cryopreservation of dried axillary buds from plantlets of Asparagus officinalis L. grown in vitro.

    Science.gov (United States)

    Uragami, A; Sakai, A; Nagai, M

    1990-10-01

    Dried axillary buds from plantlets of Asparagus lofficinalis L. grown in vitro were successfully cryopreserved. Single node segments (5mm in length) with axillary bud were taken from mature in vitro plantlets. The segments were precultured on solidfied Murashige-Skoog medium (1962) containing 0.7M sucrose at 25 °C in light for 2 days. Thereafter, these precultured segments were subjected to dehydration with silica gel at room temperature for 0 to 24 h. The axillary buds of precultured segments tolerated dehydration to about 14% water content(FW) with 50% lethality (LD50) and the threshold water content at which the dried buds remained alive after exposure to liquid nitrogen was 16.9%(LD50). The maximum rate of survival of cryopreserved buds was about 71% of untreated control. Surviving buds produced shoots and regenerated into plantlets. These results demonstrate the feasibility of cryopreserving dried axillary buds from in vitro plantlets.

  2. A Conserved Carbon Starvation Response Underlies Bud Dormancy in Woody and Herbaceous Species

    Directory of Open Access Journals (Sweden)

    Carlos Tarancón

    2017-05-01

    Full Text Available Plant shoot systems give rise to characteristic above-ground plant architectures. Shoots are formed from axillary meristems and buds, whose growth and development is modulated by systemic and local signals. These cues convey information about nutrient and water availability, light quality, sink/source organ activity and other variables that determine the timeliness and competence to maintain development of new shoots. This information is translated into a local response, in meristems and buds, of growth or quiescence. Although some key genes involved in the onset of bud latency have been identified, the gene regulatory networks (GRNs controlled by these genes are not well defined. Moreover, it has not been determined whether bud dormancy induced by environmental cues, such as a low red-to-far-red light ratio, shares genetic mechanisms with bud latency induced by other causes, such as apical dominance or a short-day photoperiod. Furthermore, the evolution and conservation of these GRNs throughout angiosperms is not well established. We have reanalyzed public transcriptomic datasets that compare quiescent and active axillary buds of Arabidopsis, with datasets of axillary buds of the woody species Vitis vinifera (grapevine and apical buds of Populus tremula x Populus alba (poplar during the bud growth-to-dormancy transition. Our aim was to identify potentially common GRNs induced during the process that leads to bud para-, eco- and endodormancy. In Arabidopsis buds that are entering eco- or paradormancy, we have identified four induced interrelated GRNs that correspond to a carbon (C starvation syndrome, typical of tissues undergoing low C supply. This response is also detectable in poplar and grapevine buds before and during the transition to dormancy. In all eukaryotes, C-limiting conditions are coupled to growth arrest and latency like that observed in dormant axillary buds. Bud dormancy might thus be partly a consequence of the underlying C

  3. Tumor budding is a strong and reproducible prognostic marker in T3N0 colorectal cancer.

    LENUS (Irish Health Repository)

    Wang, Lai Mun

    2012-02-01

    BACKGROUND: Tumor budding along the advancing front of colorectal adenocarcinoma is an early event in the metastatic process. A reproducible, prognostic budding scoring system based on outcomes in early stage colorectal cancer has not been established. DESIGN: One hundred twenty-eight T3N0M0 colorectal carcinoma patients with known outcome were identified. Tumor budding was defined as isolated tumor cells or clusters of <5 cells at the invasive tumor front. Tumor bud counts were generated in 5 regions at 200x by 2 pathologists (conventional bud count method). The median bud count per case was used to divide cases into low (median=0) and high budding (median > or =1) groups. Forty cases were reevaluated to assess reproducibility using the conventional and a novel rapid bud count method. RESULTS: Fifty-seven (45%) carcinomas had high and 71 (55%) had low budding scores. High budding was associated with an infiltrative growth pattern (P<0.0001) and lymphovascular invasion (P=0.005). Five-year cancer-specific survival was significantly poorer in high compared with low budding groups: 63% versus 91%, respectively, P<0.0001. Multivariate analysis demonstrated tumor budding to be independently prognostic (hazard ratio=4.76, P<0.001). Interobserver agreement was at least equivalent comparing the conventional to the rapid bud count methods: 87.5% agreement (kappa=0.75) versus 92.5% agreement (kappa=0.85), respectively. CONCLUSIONS: Tumor budding is a strong, reproducible, and independent prognostic marker of outcome that is easily assessed on hematoxylin and eosin slides. This may be useful for identifying the subset of T3N0M0 patients at high risk of recurrence who may benefit from adjuvant therapy.

  4. Newly identified prions in budding yeast, and their possible functions

    OpenAIRE

    Crow, Emily T.; Li, Liming

    2011-01-01

    Yeast prions are atypical genetic elements that are transmitted as heritable protein conformations. [PSI+], [URE3], and [PIN+] are three well-studied prions in the budding yeast, Saccharomyces cerevisiae. In the last three years, several additional prions have been reported in yeast, including [SWI+], [OCT+], [MCA], [GAR+], [MOT3+], [ISP+], and [NSI+]. The growing number of yeast prions suggests that protein-based inheritance might be a widespread biological phenomenon. In this review, we sum...

  5. The cell surface mucin podocalyxin regulates collective breast tumor budding.

    Science.gov (United States)

    Graves, Marcia L; Cipollone, Jane A; Austin, Pamela; Bell, Erin M; Nielsen, Julie S; Gilks, C Blake; McNagny, Kelly M; Roskelley, Calvin D

    2016-01-22

    Overexpression of the transmembrane sialomucin podocalyxin, which is known to play a role in lumen formation during polarized epithelial morphogenesis, is an independent indicator of poor prognosis in a number of epithelial cancers, including those that arise in the breast. Therefore, we set out to determine if podocalyxin plays a functional role in breast tumor progression. MCF-7 breast cancer cells, which express little endogenous podocalyxin, were stably transfected with wild type podocalyxin for forced overexpression. 4T1 mammary tumor cells, which express considerable endogenous podocalyxin, were retrovirally transduced with a short hairpin ribonucleic acid (shRNA) targeting podocalyxin for stable knockdown. In vitro, the effects of podocalyxin on collective cellular migration and invasion were assessed in two-dimensional monolayer and three-dimensional basement membrane/collagen gel culture, respectively. In vivo, local invasion was assessed after orthotopic transplantation in immunocompromised mice. Forced overexpression of podocalyxin caused cohesive clusters of epithelial MCF-7 breast tumor cells to bud off from the primary tumor and collectively invade the stroma of the mouse mammary gland in vivo. This budding was not associated with any obvious changes in histoarchitecture, matrix deposition or proliferation in the primary tumour. In vitro, podocalyxin overexpression induced a collective migration of MCF-7 tumor cells in two-dimensional (2-D) monolayer culture that was dependent on the activity of the actin scaffolding protein ezrin, a cytoplasmic binding partner of podocalyxin. In three-dimensional (3-D) culture, podocalyxin overexpression induced a collective budding and invasion that was dependent on actomyosin contractility. Interestingly, the collectively invasive cell aggregates often contained expanded microlumens that were also observed in vivo. Conversely, when endogenous podocalyxin was removed from highly metastatic, but cohesive, 4T1 mammary

  6. Saponins from the flower buds of Buddleja officinalis.

    Science.gov (United States)

    Guo, Hongzhu; Koike, Kazuo; Li, Wei; Satou, Tadaaki; Guo, Dean; Nikaido, Tamotsu

    2004-01-01

    Five new saponins, mimengosides C-G (1-5), were isolated from the flower buds of Buddleja officinalis along with five known compounds, namely, songaroside A, acteoside, phenylethyl 2-glucoside, echinacoside, and phenylethyl alcohol 8-O-beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranoside. The structures of 1-5 were elucidated using spectroscopic and chemical methods, and these compounds were evaluated for their inhibitory effects against HL-60 leukemia cells.

  7. Ecological conditions favoring budding in colonial organisms under environmental disturbance.

    Directory of Open Access Journals (Sweden)

    Mayuko Nakamaru

    Full Text Available Dispersal is a topic of great interest in ecology. Many organisms adopt one of two distinct dispersal tactics at reproduction: the production of small offspring that can disperse over long distances (such as seeds and spawned eggs, or budding. The latter is observed in some colonial organisms, such as clonal plants, corals and ants, in which (superorganisms split their body into components of relatively large size that disperse to a short distance. Contrary to the common dispersal viewpoint, short-dispersal colonial organisms often flourish even in environments with frequent disturbances. In this paper, we investigate the conditions that favor budding over long-distance dispersal of small offspring, focusing on the life history of the colony growth and the colony division ratio. These conditions are the relatively high mortality of very small colonies, logistic growth, the ability of dispersers to peacefully seek and settle unoccupied spaces, and small spatial scale of environmental disturbance. If these conditions hold, budding is advantageous even when environmental disturbance is frequent. These results suggest that the demography or life history of the colony underlies the behaviors of the colonial organisms.

  8. Trichomes control flower bud shape by linking together young petals.

    Science.gov (United States)

    Tan, Jiafu; Walford, Sally-Anne; Dennis, Elizabeth S; Llewellyn, Danny

    2016-06-20

    Trichomes are widespread in plants and develop from surface cells on different tissues(1). They have many forms and functions, from defensive spines to physical barriers that trap layers of air to insulate against desiccation, but there is growing evidence that trichomes can also have developmental roles in regulating flower structure(2,3). We report here that the trichomes on petals of cotton, Gossypium hirsutum L., are essential for correct flower bud shape through a mechanical entanglement of the trichomes on adjacent petals that anchor the edges to counter the opposing force generated by asymmetric expansion of overlapping petals. Silencing a master regulator of petal trichomes, GhMYB-MIXTA-Like10 (GhMYBML10), by RNA interference (RNAi) suppressed petal trichome growth and resulted in flower buds forming into abnormal corkscrew shapes that exposed developing anthers and stigmas to desiccation damage. Artificially gluing petal edges together could partially restore correct bud shape and fertility. Such petal 'Velcro' is present in other Malvaceae and perhaps more broadly in other plant families, although it is not ubiquitous. This mechanism for physical association between separate organs to regulate flower shape and function is different from the usual organ shape control(4) exerted through cell-to-cell communication and differential cell expansion within floral tissues(5,6).

  9. The Inside-Out Mechanism of Dicers from Budding Yeasts

    Energy Technology Data Exchange (ETDEWEB)

    D Weinberg; K Nakanishi; D Patel; D Bartel

    2011-12-31

    The Dicer ribonuclease III (RNase III) enzymes process long double-stranded RNA (dsRNA) into small interfering RNAs (siRNAs) that direct RNA interference. Here, we describe the structure and activity of a catalytically active fragment of Kluyveromyces polysporus Dcr1, which represents the noncanonical Dicers found in budding yeasts. The crystal structure revealed a homodimer resembling that of bacterial RNase III but extended by a unique N-terminal domain, and it identified additional catalytic residues conserved throughout eukaryotic RNase III enzymes. Biochemical analyses showed that Dcr1 dimers bind cooperatively along the dsRNA substrate such that the distance between consecutive active sites determines the length of the siRNA products. Thus, unlike canonical Dicers, which successively remove siRNA duplexes from the dsRNA termini, budding-yeast Dicers initiate processing in the interior and work outward. The distinct mechanism of budding-yeast Dicers establishes a paradigm for natural molecular rulers and imparts substrate preferences with ramifications for biological function.

  10. The Inside-Out Mechanism of Dicers from Budding Yeasts

    Energy Technology Data Exchange (ETDEWEB)

    Weinberg, David E.; Nakanishi, Kotaro; Patel, Dinshaw J.; Bartel, David P. (Whitehead); (MSKCC)

    2011-09-20

    The Dicer ribonuclease III (RNase III) enzymes process long double-stranded RNA (dsRNA) into small interfering RNAs (siRNAs) that direct RNA interference. Here, we describe the structure and activity of a catalytically active fragment of Kluyveromyces polysporus Dcr1, which represents the noncanonical Dicers found in budding yeasts. The crystal structure revealed a homodimer resembling that of bacterial RNase III but extended by a unique N-terminal domain, and it identified additional catalytic residues conserved throughout eukaryotic RNase III enzymes. Biochemical analyses showed that Dcr1 dimers bind cooperatively along the dsRNA substrate such that the distance between consecutive active sites determines the length of the siRNA products. Thus, unlike canonical Dicers, which successively remove siRNA duplexes from the dsRNA termini, budding-yeast Dicers initiate processing in the interior and work outward. The distinct mechanism of budding-yeast Dicers establishes a paradigm for natural molecular rulers and imparts substrate preferences with ramifications for biological function.

  11. Bud dormancy in apple trees after thermal fluctuations

    Directory of Open Access Journals (Sweden)

    Rafael Anzanello

    2014-06-01

    Full Text Available The objective of this work was to evaluate the effect of heat waves on the evolution of bud dormancy, in apple trees with contrasting chilling requirements. Twigs of 'Castel Gala' and 'Royal Gala' were collected in orchards in Papanduva, state of Santa Catarina, Brazil, and were exposed to constant (3°C or alternating (3 and 15°C for 12/12 hours temperature, combined with zero, one or two days a week at 25°C. Two additional treatments were evaluated: constant temperature (3°C, with a heat wave of seven days at 25°C, in the beginning or in the middle of the experimental period. Periodically, part of the twigs was transferred to 25°C for daily budburst evaluation of apical and lateral buds. Endodormancy (dormancy induced by cold was overcome with less than 330 chilling hours (CH of constant cold in 'Castel Gala' and less than 618 CH in 'Royal Gala'. A daily 15°C-temperature cycle did not affect the endodormancy process. Heat waves during endodormancy resulted in an increased CH to achieve bud requirements. The negative effect of high temperature depended on the lasting of this condition. Chilling was partly cancelled during dormancy when the heat wave lasted 36 continuous hours or more. Therefore, budburst prediction models need adjustments, mainly for regions with mild and irregular winters, such as those of Southern Brazil.

  12. Epithelial to mesenchymal transition correlates with tumor budding and predicts prognosis in esophageal squamous cell carcinoma.

    Science.gov (United States)

    Niwa, Yukiko; Yamada, Suguru; Koike, Masahiko; Kanda, Mitsuro; Fujii, Tsutomu; Nakayama, Goro; Sugimoto, Hiroyuki; Nomoto, Shuji; Fujiwara, Michitaka; Kodera, Yasuhiro

    2014-11-01

    Epithelial to mesenchymal transition (EMT) is considered to play an important role in cancer invasion. Tumor budding is a prognostic factor in esophageal squamous cell carcinoma (ESCC). The aim of this study was to explore the correlation between EMT and tumor budding. Surgical specimens from 78 cases of ESCC resected without preoperative treatment between 2001 and 2013 were enrolled in the study. The mRNA expressions of E-cadherin and vimentin were measured in cancerous tissues using real-time PCR, and each tumor was classified into either epithelial or mesenchymal group. Tumor budding was evaluated in H&E-stained slides and divided into two groups; low-grade budding (budding (≥3). The 5-year survival rate in the epithelial group was significantly higher than that in the mesenchymal group (62.0% vs. 31.5%, P = 0.021). Survival rate of patients in the low-grade budding group was significantly higher than that of patients in the high-grade budding group (75.1% vs. 25.9%, P tumor budding was significantly associated with the mesenchymal group (P = 0.009). EMT was found to occur in ESCC and was significantly associated with tumor budding. Tumor budding was identified as a significant independent prognostic factor among the current population of ESCC. © 2014 Wiley Periodicals, Inc.

  13. Ice nucleation activity in various tissues of Rhododendron flower buds: their relevance to extraorgan freezing

    Directory of Open Access Journals (Sweden)

    Masaya eIshikawa

    2015-03-01

    Full Text Available Wintering flower buds of cold hardy Rhododendron japonicum cooled slowly to subfreezing temperatures are known to undergo extraorgan freezing, whose mechanisms remain obscure. We revisited this material to demonstrate why bud scales freeze first in spite of their lower water content, why florets remain deeply supercooled and how seasonal adaptive responses occur in regard to extraorgan freezing in flower buds. We determined ice nucleation activity (INA of various flower bud tissues of using a test tube-based assay. Irrespective of collection sites, outer and inner bud scales that function as ice sinks in extraorgan freezing had high INA levels whilst florets that remain supercooled and act as a water source lacked INA. The INA level of bud scales was not high in late August when flower bud formation was ending, but increased to reach the highest level in late October just before the first autumnal freeze. The results support the following hypothesis: the high INA in bud scales functions as the subfreezing sensor, ensuring the primary freezing in bud scales at warmer subzero temperatures, which likely allows the migration of floret water to the bud scales and accumulation of icicles within the bud scales. The low INA in the florets helps them remain unfrozen by deep supercooling. The INA in the bud scales was resistant to grinding and autoclaving at 121°C for 15 min, implying the intrinsic nature of the INA rather than of microbial origin, whilst the INA in stem bark was autoclaving labile. Anti-nucleation activity (ANA was implicated in the leachate of autoclaved bud scales, which suppresses the INA at millimolar levels of concentration and likely differs from the colligative effects of the solutes. The tissue INA levels likely contribute to the establishment of freezing behaviors by ensuring the order of freezing in the tissues: from the primary freeze to the last tissue remaining unfrozen.

  14. Ice nucleation activity in various tissues of Rhododendron flower buds: their relevance to extraorgan freezing.

    Science.gov (United States)

    Ishikawa, Masaya; Ishikawa, Mikiko; Toyomasu, Takayuki; Aoki, Takayuki; Price, William S

    2015-01-01

    Wintering flower buds of cold hardy Rhododendron japonicum cooled slowly to subfreezing temperatures are known to undergo extraorgan freezing, whose mechanisms remain obscure. We revisited this material to demonstrate why bud scales freeze first in spite of their lower water content, why florets remain deeply supercooled and how seasonal adaptive responses occur in regard to extraorgan freezing in flower buds. We determined ice nucleation activity (INA) of various flower bud tissues using a test tube-based assay. Irrespective of collection sites, outer and inner bud scales that function as ice sinks in extraorgan freezing had high INA levels whilst florets that remain supercooled and act as a water source lacked INA. The INA level of bud scales was not high in late August when flower bud formation was ending, but increased to reach the highest level in late October just before the first autumnal freeze. The results support the following hypothesis: the high INA in bud scales functions as the subfreezing sensor, ensuring the primary freezing in bud scales at warmer subzero temperatures, which likely allows the migration of floret water to the bud scales and accumulation of icicles within the bud scales. The low INA in the florets helps them remain unfrozen by deep supercooling. The INA in the bud scales was resistant to grinding and autoclaving at 121(∘)C for 15 min, implying the intrinsic nature of the INA rather than of microbial origin, whilst the INA in stem bark was autoclaving-labile. Anti-nucleation activity (ANA) was implicated in the leachate of autoclaved bud scales, which suppresses the INA at millimolar levels of concentration and likely differs from the colligative effects of the solutes. The tissue INA levels likely contribute to the establishment of freezing behaviors by ensuring the order of freezing in the tissues: from the primary freeze to the last tissue remaining unfrozen.

  15. Tumor budding, a novel prognostic indicator for predicting stage progression in T1 bladder cancers.

    Science.gov (United States)

    Fukumoto, Keishiro; Kikuchi, Eiji; Mikami, Shuji; Ogihara, Koichiro; Matsumoto, Kazuhiro; Miyajima, Akira; Oya, Mototsugu

    2016-09-01

    Tumor budding has been defined as an isolated single cancer cell or a cluster composed of fewer than five cancer cells scattered in the stroma. It is a strong predictor for lymph node metastasis in T1 colorectal cancer. We introduced this concept to T1 non-muscle invasive bladder cancer and evaluated whether tumor budding could have a prognostic impact on the clinical outcome. We identified 121 consecutive patients with newly diagnosed T1 bladder cancer between 1994 and 2014 at Keio University Hospital. All slides were re-reviewed by a dedicated uropathologist. Budding foci were counted under ×200 magnification. When the number of budding foci was 10 or more, tumor budding was defined as positive. The relationship between tumor budding and clinical outcomes was assessed using a multivariate analysis. The median follow-up was 52 months. Tumor budding was positive in 21 patients (17.4%). Tumor budding was significantly associated with T1 substaging, tumor architecture and lymphovascular invasion. The 5-year progression-free survival rate in T1 bladder cancer patients with tumor budding was 53.8%, which was significantly lower than that in patients without tumor budding (88.4%, P = 0.001). A multivariate Cox regression analysis revealed that tumor budding was independently associated with stage progression (P = 0.002, hazard ratio = 4.90). In a subgroup of patients treated with bacillus Calmette-Guérin instillation (n = 88), tumor budding was also independently associated with stage progression (P = 0.003, hazard ratio = 5.65). Tumor budding may be a novel indicator for predicting stage progression in T1 bladder cancer, and would likely be easily introduced in clinical practice. © 2016 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.

  16. Tumor Budding as a Strong Prognostic Indicator in Invasive Ampullary Adenocarcinomas

    Science.gov (United States)

    Ohike, Nobuyuki; Coban, Ipek; Kim, Grace E.; Basturk, Olca; Tajiri, Takuma; Krasinskas, Alyssa; Bandyopadhyay, Sudeshna; Morohoshi, Toshio; Shimada, Yuki; Kooby, David A.; Staley, Charles A.; Goodman, Michael; Adsay, Nazmi Volkan

    2011-01-01

    Prognostication of invasive ampullary adenocarcinomas (AACs) and their stratification into appropriate management categories have been highly challenging owing to a lack of well-established predictive parameters. In colorectal cancers, recent studies have shown that tumor budding confers a worse prognosis and correlates significantly with nodal metastasis and recurrence; however, this has not been evaluated in AAC. To investigate the prevalence, significance, and clinical correlations of tumor budding in AAC, 244 surgically resected, stringently defined, invasive AAC were analyzed for tumor budding—defined as the presence of more than or equal to 5 isolated single cancer cells or clusters composed of fewer than 5 cancer cells per field measuring 0.785 mm2 using a 20 × objective lens in the stroma of the invasive front. The extent of the budding was then further classified as “high” if there were greater than or equal to 3 budding foci and as “low” if there were budding foci or no budding focus. One hundred ninety-four AACs (80%) were found to be high-budding and 50 (20%) were low-budding. When the clinicopathologic features and survival of the 2 groups were compared, the AACs with high-budding had larger invasion size (19 mm vs. 13 mm; Ptumor budding was found to be an independent predictor of survival (P = 0.01), which impacts prognosis (hazard ratio: 2.6) even more than T-stage and lymph node metastasis (hazard ratio: 1.9 and 1.8, respectively). In conclusion, tumor budding is frequently encountered in AAC. High-budding is a strong independent predictor of overall survival, with a prognostic correlation stronger than the 2 established parameters: T-stage and lymph node metastasis. Therefore, budding should be incorporated into surgical pathology reports for AAC. PMID:20871215

  17. Involvement of EARLY BUD-BREAK, an AP2/ERF Transcription Factor Gene, in Bud Break in Japanese Pear (Pyrus pyrifolia Nakai) Lateral Flower Buds: Expression, Histone Modifications and Possible Target Genes.

    Science.gov (United States)

    Anh Tuan, Pham; Bai, Songling; Saito, Takanori; Imai, Tsuyoshi; Ito, Akiko; Moriguchi, Takaya

    2016-05-01

    In the Japanese pear (Pyrus pyrifolia Nakai) 'Kosui', three developmental stages of lateral flower buds have been proposed to occur during ecodormancy to the flowering phase, i.e. rapid enlargement, sprouting and flowering. Here, we report an APETALA2/ethylene-responsive factor (AP2/ERF) transcription factor gene, named pear EARLY BUD-BREAK (PpEBB), which was highly expressed during the rapid enlargement stage occurring prior to the onset of bud break in flower buds. Gene expression analysis revealed that PpEBB expression was dramatically increased during the rapid enlargement stage in three successive growing seasons. PpEBB transcript levels peaked 1 week prior to onset of bud break in 'Kosui' potted plants treated with hydrogen cyanamide or water under forcing conditions. Chromatin immunoprecipitation-quantitative PCR showed that higher levels of active histone modifications (trimethylation of the histone H3 tail at Lys4) in the 5'-upstream and start codon regions of the PpEBB gene were associated with the induced expression level of PpEBB during the rapid enlargement stage. In addition, we provide evidence that PpEBB may interact with and regulate pear four D-type cyclin (PpCYCD3) genes during bud break in 'Kosui' lateral flower buds. PpEBB significantly increased the promoter activities of four PpCYCD3 genes in a dual-luciferase assay using tobacco leaves. Taken together, our findings uncovered aspects of the bud break regulatory mechanism in the Japanese pear and provided further evidence that the EBB family plays an important role in bud break in perennial plants. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  18. Sucrose promotes axillary bud outgrowth in Rosa hybrida and plays a signal role during this process

    OpenAIRE

    Barbier, François; Bertheloot, Jessica; Peron, Thomas; Perez-Garcia, Maria Dolores; Sakr, Soulaiman

    2013-01-01

    Shoot branching is a developmental process by which axillary buds are released from dormancy and develop into new axes. Bud outgrowth is largely affected by a number of environmental factors such as temperature, air and soil humidity, gravitropism and light, confering thus plasticity to the plant development. Control of bud outgrowth is thereby a key mechanism in the establishment of plant architecture in response to environment. Sugars, whose levels in plant are also highly dependent on the ...

  19. Phenology of perennial, native grass, belowground axillary buds in the northern mixed-grass prairie.

    Science.gov (United States)

    Russell, Morgan L; Vermeire, Lance T; Ganguli, Amy C; Hendrickson, John R

    2017-06-01

    Vegetative reproduction from belowground bud banks is the primary driver of grassland systems. Despite the importance of bud banks, the timing of recruitment and the crucial link between formation and maintenance is unknown. We assessed patterns of belowground bud development, dormancy, and mortality associated with three perennial native grasses in the northern Great Plains. Temperature and soil moisture were measured below the soil surface to determine relationships with belowground bud development. Blue grama (Bouteloua gracilis) generated more buds over winter that remained dormant; whereas, C3 species needle-and-thread (Hesperostipa comata) and western wheatgrass (Pascopyrum smithii), maintained limited dormant buds throughout winter. Soil temperature was a good predictor for C4 species bud production; whereas, soil moisture was a reliable predictor for C3 buds. Distinct differences existed between C4 species blue grama and C3 species needle-and-thread, whereas C3 species western wheatgrass (Pascopyrum smithii) was intermediate, indicating there is likely a species-specific continuum between the C3 and C4 extremes rather than a stark difference. The ability to predict belowground bud development is a novel insight to native perennial grasses. Native grass species' strategies and adaptability regarding belowground bud bank size and bud phenology are important factors optimizing tiller recruitment given the variable growing conditions. Patterns of bud dormancy and development will provide insight to the underlying mechanisms by which management practices and fluctuations in precipitation amount and growing season length can alter mixed-grass prairie plant community dynamics. © 2017 Botanical Society of America.

  20. The bud rot of oil palm in San Lorenzo, Esmeraldas province, Ecuador

    OpenAIRE

    Fernando Rivas Figueroa; Lidcay Herrera Isla

    2015-01-01

    Oil palms (Elaeis guineensis Jacq) in the area of San Lorenzo were directly observed, and some plants were dissected to assess the internal symptoms, with the purpose of characterizing the symptomatology of bud rot. The plants showed chlorosis and yellowing of young leaves around the bud, necrosis of leaflets in young leaves, necrosis and rot of spears (outer leaf in the process of opening), bending of spears leaves due to the breaking in the lower third, necrosis and internal bud decay which...

  1. Sucrose is an early modulator of the key hormonal mechanisms controlling bud outgrowth in Rosa hybrida.

    Science.gov (United States)

    Barbier, François; Péron, Thomas; Lecerf, Marion; Perez-Garcia, Maria-Dolores; Barrière, Quentin; Rolčík, Jakub; Boutet-Mercey, Stéphanie; Citerne, Sylvie; Lemoine, Remi; Porcheron, Benoît; Roman, Hanaé; Leduc, Nathalie; Le Gourrierec, José; Bertheloot, Jessica; Sakr, Soulaiman

    2015-05-01

    Sugar has only recently been identified as a key player in triggering bud outgrowth, while hormonal control of bud outgrowth is already well established. To get a better understanding of sugar control, the present study investigated how sugar availability modulates the hormonal network during bud outgrowth in Rosa hybrida. Other plant models, for which mutants are available, were used when necessary. Buds were grown in vitro to manipulate available sugars. The temporal patterns of the hormonal regulatory network were assessed in parallel with bud outgrowth dynamics. Sucrose determined bud entrance into sustained growth in a concentration-dependent manner. Sustained growth was accompanied by sustained auxin production in buds, and sustained auxin export in a DR5::GUS-expressing pea line. Several events occurred ahead of sucrose-stimulated bud outgrowth. Sucrose upregulated early auxin synthesis genes (RhTAR1, RhYUC1) and the auxin efflux carrier gene RhPIN1, and promoted PIN1 abundance at the plasma membrane in a pPIN1::PIN1-GFP-expressing tomato line. Sucrose downregulated both RwMAX2, involved in the strigolactone-transduction pathway, and RhBRC1, a repressor of branching, at an early stage. The presence of sucrose also increased stem cytokinin content, but sucrose-promoted bud outgrowth was not related to that pathway. In these processes, several non-metabolizable sucrose analogues induced sustained bud outgrowth in R. hybrida, Pisum sativum, and Arabidopsis thaliana, suggesting that sucrose was involved in a signalling pathway. In conclusion, we identified potential hormonal candidates for bud outgrowth control by sugar. They are central to future investigations aimed at disentangling the processes that underlie regulation of bud outgrowth by sugar. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  2. Relationship between formation of gametophore buds in the protonema of mosses and increase in ribonuclease activity

    Directory of Open Access Journals (Sweden)

    M. Spychała

    2015-01-01

    Full Text Available Changes in RNase activity similar to those accompanying cytokinin-induced formation of gametophore buds in mosses (a decrease in the early phase of bud formation and later an increase in enzyme activity have also been found during spontaneous formation of gametophores in moss ontogenesis. Using various factors affecting the cytokinin-induced process of bud formation a correlation has been found between this process and the increase in RNase activity.

  3. Spaceflight enhances cell aggregation and random budding in Candida albicans.

    Directory of Open Access Journals (Sweden)

    Aurélie Crabbé

    Full Text Available This study presents the first global transcriptional profiling and phenotypic characterization of the major human opportunistic fungal pathogen, Candida albicans, grown in spaceflight conditions. Microarray analysis revealed that C. albicans subjected to short-term spaceflight culture differentially regulated 452 genes compared to synchronous ground controls, which represented 8.3% of the analyzed ORFs. Spaceflight-cultured C. albicans-induced genes involved in cell aggregation (similar to flocculation, which was validated by microscopic and flow cytometry analysis. We also observed enhanced random budding of spaceflight-cultured cells as opposed to bipolar budding patterns for ground samples, in accordance with the gene expression data. Furthermore, genes involved in antifungal agent and stress resistance were differentially regulated in spaceflight, including induction of ABC transporters and members of the major facilitator family, downregulation of ergosterol-encoding genes, and upregulation of genes involved in oxidative stress resistance. Finally, downregulation of genes involved in actin cytoskeleton was observed. Interestingly, the transcriptional regulator Cap1 and over 30% of the Cap1 regulon was differentially expressed in spaceflight-cultured C. albicans. A potential role for Cap1 in the spaceflight response of C. albicans is suggested, as this regulator is involved in random budding, cell aggregation, and oxidative stress resistance; all related to observed spaceflight-associated changes of C. albicans. While culture of C. albicans in microgravity potentiates a global change in gene expression that could induce a virulence-related phenotype, no increased virulence in a murine intraperitoneal (i.p. infection model was observed under the conditions of this study. Collectively, our data represent an important basis for the assessment of the risk that commensal flora could play during human spaceflight missions. Furthermore, since the

  4. Metabolite changes in conifer buds and needles during forced bud break in Norway spruce (Picea abies and European silver fir (Abies alba

    Directory of Open Access Journals (Sweden)

    Priyanka eDhuli

    2014-12-01

    Full Text Available Environmental changes such as early spring and warm spells induce bud burst and photosynthetic processes in cold-acclimated coniferous trees and consequently, cellular metabolism in overwintering needles and buds. The purpose of the study was to examine metabolism in conifers under forced deacclimation (artificially induced spring by exposing shoots of Picea abies (boreal species and Abies alba (temperate species to a greenhouse environment (22°C, 16/8 h D/N cycle over a nine week period. Each week, we scored bud opening and collected samples for GC/MS–based metabolite profiling. We detected a total of 169 assigned metabolites and 80 identified metabolites, comprising compounds such as mono- and disaccharides, Krebs cycle acids, amino acids, polyols, phenolics and phosphorylated structures. Untargeted multivariate statistical analysis based on PCA and cluster analysis segregated samples by species, tissue type, and stage of tissue deacclimations. Similar patterns of metabolic regulation in both species were observed in buds (amino acids, Krebs cycle acids and needles (hexoses, pentoses, and Krebs cycle acids. Based on correlation of bud opening score with compound levels, distinct metabolites could be associated with bud and shoot development, including amino acids, sugars and acids with known osmolyte function, and secondary metabolites. This study has shed light on how elevated temperature affects metabolism in buds and needles of conifer species during the deacclimation phase, and contributes to the discussion about how phenological characters in conifers may respond to future global warming.

  5. β-Catenin signaling regulates temporally discrete phases of anterior taste bud development

    Science.gov (United States)

    Thirumangalathu, Shoba; Barlow, Linda A.

    2015-01-01

    The sense of taste is mediated by multicellular taste buds located within taste papillae on the tongue. In mice, individual taste buds reside in fungiform papillae, which develop at mid-gestation as epithelial placodes in the anterior tongue. Taste placodes comprise taste bud precursor cells, which express the secreted factor sonic hedgehog (Shh) and give rise to taste bud cells that differentiate around birth. We showed previously that epithelial activation of β-catenin is the primary inductive signal for taste placode formation, followed by taste papilla morphogenesis and taste bud differentiation, but the degree to which these later elements were direct or indirect consequences of β-catenin signaling was not explored. Here, we define discrete spatiotemporal functions of β-catenin in fungiform taste bud development. Specifically, we show that early epithelial activation of β-catenin, before taste placodes form, diverts lingual epithelial cells from a taste bud fate. By contrast, β-catenin activation a day later within Shh+ placodes, expands taste bud precursors directly, but enlarges papillae indirectly. Further, placodal activation of β-catenin drives precocious differentiation of Type I glial-like taste cells, but not other taste cell types. Later activation of β-catenin within Shh+ precursors during papilla morphogenesis also expands taste bud precursors and accelerates Type I cell differentiation, but papilla size is no longer enhanced. Finally, although Shh regulates taste placode patterning, we find that it is dispensable for the accelerated Type I cell differentiation induced by β-catenin. PMID:26525674

  6. Chemical composition of volatile oil from Cinnamomum zeylanicum buds.

    Science.gov (United States)

    Jayaprakasha, Guddadarangavvanahally K; Rao, Lingamallu Jaganmohan; Sakariah, Kunnumpurath K

    2002-01-01

    The hydro-distilled volatile oil of the Cinnamomum zeylanicum (C. zeylanicum) buds was analyzed using GC and GC-MS for the first time. Thirty-four compounds representing approximately 98% of the oil was characterized. It consists of terpene hydrocarbons (78%) and oxygenated terpenoids (9%). alpha-Bergamotene (27.38%) and alpha-copaene (23.05%) are found to be the major compounds. A comparison of the chemical composition of the oil was made with that of flowers and fruits.

  7. UXO Detection and Characterization using new Berkeley UXO Discriminator (BUD)

    Science.gov (United States)

    Gasperikova, E.; Morrison, H. F.; Smith, J. T.; Becker, A.

    2006-05-01

    An optimally designed active electromagnetic system (AEM), Berkeley UXO Discriminator, BUD, has been developed for detection and characterization of UXO in the 20 mm to 150 mm size range. The system incorporates three orthogonal transmitters, and eight pairs of differenced receivers. The transmitter-receiver assembly together with the acquisition box, as well as the battery power and GPS receiver, is mounted on a small cart to assure system mobility. BUD not only detects the object itself but also quantitatively determines its size, shape, orientation, and metal content (ferrous or non-ferrous, mixed metals). Moreover, the principal polarizabilities and size of a metallic target can be determined from a single position of the BUD platform. The search for UXO is a two-step process. The object must first be detected and its location determined then the parameters of the object must be defined. A satisfactory classification scheme is one that determines the principal dipole polarizabilities of a target. While UXO objects have a single major polarizability (principal moment) coincident with the long axis of the object and two equal transverse polarizabilities, the scrap metal has all three principal moments entirely different. This description of the inherent polarizabilities of a target is a major advance in discriminating UXO from irregular scrap metal. Our results clearly show that BUD can resolve the intrinsic polarizabilities of a target and that there are very clear distinctions between symmetric intact UXO and irregular scrap metal. Target properties are determined by an inversion algorithm, which at any given time inverts the response to yield the location (x, y, z) of the target, its attitude and its principal polarizabilities (yielding an apparent aspect ratio). Signal-to-noise estimates (or measurements) are interpreted in this inversion to yield error estimates on the location, attitude and polarizabilities. This inversion at a succession of times provides

  8. Evolutionary biology through the lens of budding yeast comparative genomics.

    Science.gov (United States)

    Marsit, Souhir; Leducq, Jean-Baptiste; Durand, Éléonore; Marchant, Axelle; Filteau, Marie; Landry, Christian R

    2017-10-01

    The budding yeast Saccharomyces cerevisiae is a highly advanced model system for studying genetics, cell biology and systems biology. Over the past decade, the application of high-throughput sequencing technologies to this species has contributed to this yeast also becoming an important model for evolutionary genomics. Indeed, comparative genomic analyses of laboratory, wild and domesticated yeast populations are providing unprecedented detail about many of the processes that govern evolution, including long-term processes, such as reproductive isolation and speciation, and short-term processes, such as adaptation to natural and domestication-related environments.

  9. Functionally homologous DNA replication genes in fission and budding yeast

    OpenAIRE

    Sánchez, Mar; Calzada, Arturo; Bueno, Avelino

    1999-01-01

    The cdc18+ gene of the fission yeast Schizosaccharomyces pombe is involved in the initiation of DNA replication as well as in coupling the S phase to mitosis. In this work, we show that the Saccharomyces cerevisiae CDC6 gene complements cdc18-K46 ts and cdc18 deletion mutant S. pombe strains. The budding yeast gene suppresses both the initiation and the checkpoint defects associated with the lack of cdc18+. The Cdc6 protein interacts in vivo with Cdc2 kinase complexes. Interestingly, Cdc6 is ...

  10. Naumovozyma castellii: an alternative model for budding yeast molecular biology.

    Science.gov (United States)

    Karademir Andersson, Ahu; Cohn, Marita

    2017-03-01

    Naumovozyma castellii (Saccharomyces castellii) is a member of the budding yeast family Saccharomycetaceae. It has been extensively used as a model organism for telomere biology research and has gained increasing interest as a budding yeast model for functional analyses owing to its amenability to genetic modifications. Owing to the suitable phylogenetic distance to S. cerevisiae, the whole genome sequence of N. castellii has provided unique data for comparative genomic studies, and it played a key role in the establishment of the timing of the whole genome duplication and the evolutionary events that took place in the subsequent genomic evolution of the Saccharomyces lineage. Here we summarize the historical background of its establishment as a laboratory yeast species, and the development of genetic and molecular tools and strains. We review the research performed on N. castellii, focusing on areas where it has significantly contributed to the discovery of new features of molecular biology and to the advancement of our understanding of molecular evolution. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  11. Inhibition of budding/release of porcine endogenous retrovirus.

    Science.gov (United States)

    Abe, Masumi; Fukuma, Aiko; Yoshikawa, Rokusuke; Miyazawa, Takayuki; Yasuda, Jiro

    2014-08-01

    PERV is integrated into the genome of all pigs. PERV-A and PERV-B are polytropic and can productively infect human cell lines, whereas PERV-C is ecotropic. Recombinant PERV-A/C can infect human cells and exhibits high titer replication. Therefore, use of pigs for human xenotransplantation raises concerns about the risks of transfer of this infectious agent from donors to xenotransplantation recipients. To establish strategies to inhibit PERV production from cells, in the present study, we investigated the mechanism of PERV budding and anti-PERV activity of Tetherin/BST-2. The results showed that DN mutants of WWP-2, Tsg101, and Vps4A/B markedly reduced PERV production in human and porcine cell lines, suggesting that PERV budding uses these cellular factors and the cellular MVB sorting pathway as well as many other retroviruses. Moreover, PERV production was also reduced by human and porcine Tetherin/BST-2. These data are useful for developing strategies to inhibit PERV production and may reduce the risk of PERV infection in xenotransplantation. © 2014 The Societies and Wiley Publishing Asia Pty Ltd.

  12. A computational clonal analysis of the developing mouse limb bud.

    Directory of Open Access Journals (Sweden)

    Luciano Marcon

    Full Text Available A comprehensive spatio-temporal description of the tissue movements underlying organogenesis would be an extremely useful resource to developmental biology. Clonal analysis and fate mappings are popular experiments to study tissue movement during morphogenesis. Such experiments allow cell populations to be labeled at an early stage of development and to follow their spatial evolution over time. However, disentangling the cumulative effects of the multiple events responsible for the expansion of the labeled cell population is not always straightforward. To overcome this problem, we develop a novel computational method that combines accurate quantification of 2D limb bud morphologies and growth modeling to analyze mouse clonal data of early limb development. Firstly, we explore various tissue movements that match experimental limb bud shape changes. Secondly, by comparing computational clones with newly generated mouse clonal data we are able to choose and characterize the tissue movement map that better matches experimental data. Our computational analysis produces for the first time a two dimensional model of limb growth based on experimental data that can be used to better characterize limb tissue movement in space and time. The model shows that the distribution and shapes of clones can be described as a combination of anisotropic growth with isotropic cell mixing, without the need for lineage compartmentalization along the AP and PD axis. Lastly, we show that this comprehensive description can be used to reassess spatio-temporal gene regulations taking tissue movement into account and to investigate PD patterning hypothesis.

  13. Iniciation in vitro of buds of Ipomoea batatas

    Directory of Open Access Journals (Sweden)

    Orlando S. González Paneque

    2003-04-01

    Full Text Available Different buds of sweet potatowere studied. Roots of the clones Cemsa 78-354, Inivit 90-1, Inivit 93-1, Yabú-8 and Jewel were selected and put in glass with water in the laboratories for inducing the shoots. Afterwards the top shoot and the axiles shoots one to five (from top shoot were selected and sown in the Murashige and Skoog medium with the sales MS, Thiamine (1 mg.l-1, myoinositol (100 mg.l-1, sucrose (3%, gelrite (2 g.l-1 and different growth regulator combinations of giberelic acid (AG3 and 3-indol acetic acid (AIA, for example: AG3 (5 mg.l-1 + AIA (0.05 mg.l-1, AG3 (10 mg.l-1 + AIA (0.05 mg.l-1, AG3 (5 mg.l-1 + AIA (0.1 mg.l-1 and AG3 (10 mg.l-1 + AIA (0.1 mg.l-1. The pH was adjusted to 5.8. After five weeks, the morphologic behaviuor of the shoots was evaluated. The better results were obtained in the micropropagation with the use of the axiles shoots two, three and four in the culture medium whit growth regulator AG3 (10 mg.l-1 + AIA (0.05 mg.l-1. Key words: clones, axiles buds, top shoot, brotation

  14. Origin plasticity during budding yeast DNA replication in vitro

    Science.gov (United States)

    Gros, Julien; Devbhandari, Sujan; Remus, Dirk

    2014-01-01

    The separation of DNA replication origin licensing and activation in the cell cycle is essential for genome stability across generations in eukaryotic cells. Pre-replicative complexes (pre-RCs) license origins by loading Mcm2-7 complexes in inactive form around DNA. During origin firing in S phase, replisomes assemble around the activated Mcm2-7 DNA helicase. Budding yeast pre-RCs have previously been reconstituted in vitro with purified proteins. Here, we show that reconstituted pre-RCs support replication of plasmid DNA in yeast cell extracts in a reaction that exhibits hallmarks of cellular replication initiation. Plasmid replication in vitro results in the generation of covalently closed circular daughter molecules, indicating that the system recapitulates the initiation, elongation, and termination stages of DNA replication. Unexpectedly, yeast origin DNA is not strictly required for DNA replication in vitro, as heterologous DNA sequences could support replication of plasmid molecules. Our findings support the notion that epigenetic mechanisms are important for determining replication origin sites in budding yeast, highlighting mechanistic principles of replication origin specification that are common among eukaryotes. PMID:24566988

  15. Signal transduction and information processing in mammalian taste buds

    Science.gov (United States)

    2013-01-01

    The molecular machinery for chemosensory transduction in taste buds has received considerable attention within the last decade. Consequently, we now know a great deal about sweet, bitter, and umami taste mechanisms and are gaining ground rapidly on salty and sour transduction. Sweet, bitter, and umami tastes are transduced by G-protein-coupled receptors. Salty taste may be transduced by epithelial Na channels similar to those found in renal tissues. Sour transduction appears to be initiated by intracellular acidification acting on acid-sensitive membrane proteins. Once a taste signal is generated in a taste cell, the subsequent steps involve secretion of neurotransmitters, including ATP and serotonin. It is now recognized that the cells responding to sweet, bitter, and umami taste stimuli do not possess synapses and instead secrete the neurotransmitter ATP via a novel mechanism not involving conventional vesicular exocytosis. ATP is believed to excite primary sensory afferent fibers that convey gustatory signals to the brain. In contrast, taste cells that do have synapses release serotonin in response to gustatory stimulation. The postsynaptic targets of serotonin have not yet been identified. Finally, ATP secreted from receptor cells also acts on neighboring taste cells to stimulate their release of serotonin. This suggests that there is important information processing and signal coding taking place in the mammalian taste bud after gustatory stimulation. PMID:17468883

  16. GATA6 Is a Crucial Regulator of Shh in the Limb Bud

    Science.gov (United States)

    Kozhemyakina, Elena; Ionescu, Andreia; Lassar, Andrew B.

    2014-01-01

    In the limb bud, patterning along the anterior-posterior (A-P) axis is controlled by Sonic Hedgehog (Shh), a signaling molecule secreted by the “Zone of Polarizing Activity”, an organizer tissue located in the posterior margin of the limb bud. We have found that the transcription factors GATA4 and GATA6, which are key regulators of cell identity, are expressed in an anterior to posterior gradient in the early limb bud, raising the possibility that GATA transcription factors may play an additional role in patterning this tissue. While both GATA4 and GATA6 are expressed in an A-P gradient in the forelimb buds, the hindlimb buds principally express GATA6 in an A-P gradient. Thus, to specifically examine the role of GATA6 in limb patterning we generated Prx1-Cre; GATA6fl/fl mice, which conditionally delete GATA6 from their developing limb buds. We found that these animals display ectopic expression of both Shh and its transcriptional targets specifically in the anterior mesenchyme of the hindlimb buds. Loss of GATA6 in the developing limbs results in the formation of preaxial polydactyly in the hindlimbs. Conversely, forced expression of GATA6 throughout the limb bud represses expression of Shh and results in hypomorphic limbs. We have found that GATA6 can bind to chromatin (isolated from limb buds) encoding either Shh or Gli1 regulatory elements that drive expression of these genes in this tissue, and demonstrated that GATA6 works synergistically with FOG co-factors to repress expression of luciferase reporters driven by these sequences. Most significantly, we have found that conditional loss of Shh in limb buds lacking GATA6 prevents development of hindlimb polydactyly in these compound mutant embryos, indicating that GATA6 expression in the anterior region of the limb bud blocks hindlimb polydactyly by repressing ectopic expression of Shh. PMID:24415953

  17. A fate map of the murine pancreas buds reveals a multipotent ventral foregut organ progenitor.

    Directory of Open Access Journals (Sweden)

    Jesse R Angelo

    Full Text Available The definitive endoderm is the embryonic germ layer that gives rise to the budding endodermal organs including the thyroid, lung, liver and pancreas as well as the remainder of the gut tube. DiI fate mapping and whole embryo culture were used to determine the endodermal origin of the 9.5 days post coitum (dpc dorsal and ventral pancreas buds. Our results demonstrate that the progenitors of each bud occupy distinct endodermal territories. Dorsal bud progenitors are located in the medial endoderm overlying somites 2-4 between the 2 and 11 somite stage (SS. The endoderm forming the ventral pancreas bud is found in 2 distinct regions. One territory originates from the left and right lateral endoderm caudal to the anterior intestinal portal by the 6 SS and the second domain is derived from the ventral midline of the endoderm lip (VMEL. Unlike the laterally located ventral foregut progenitors, the VMEL population harbors a multipotent progenitor that contributes to the thyroid bud, the rostral cap of the liver bud, ventral midline of the liver bud and the midline of the ventral pancreas bud in a temporally restricted manner. This data suggests that the midline of the 9.5 dpc thyroid, liver and ventral pancreas buds originates from the same progenitor population, demonstrating a developmental link between all three ventral foregut buds. Taken together, these data define the location of the dorsal and ventral pancreas progenitors in the prespecified endodermal sheet and should lead to insights into the inductive events required for pancreas specification.

  18. GATA6 is a crucial regulator of Shh in the limb bud.

    Directory of Open Access Journals (Sweden)

    Elena Kozhemyakina

    2014-01-01

    Full Text Available In the limb bud, patterning along the anterior-posterior (A-P axis is controlled by Sonic Hedgehog (Shh, a signaling molecule secreted by the "Zone of Polarizing Activity", an organizer tissue located in the posterior margin of the limb bud. We have found that the transcription factors GATA4 and GATA6, which are key regulators of cell identity, are expressed in an anterior to posterior gradient in the early limb bud, raising the possibility that GATA transcription factors may play an additional role in patterning this tissue. While both GATA4 and GATA6 are expressed in an A-P gradient in the forelimb buds, the hindlimb buds principally express GATA6 in an A-P gradient. Thus, to specifically examine the role of GATA6 in limb patterning we generated Prx1-Cre; GATA6(fl/fl mice, which conditionally delete GATA6 from their developing limb buds. We found that these animals display ectopic expression of both Shh and its transcriptional targets specifically in the anterior mesenchyme of the hindlimb buds. Loss of GATA6 in the developing limbs results in the formation of preaxial polydactyly in the hindlimbs. Conversely, forced expression of GATA6 throughout the limb bud represses expression of Shh and results in hypomorphic limbs. We have found that GATA6 can bind to chromatin (isolated from limb buds encoding either Shh or Gli1 regulatory elements that drive expression of these genes in this tissue, and demonstrated that GATA6 works synergistically with FOG co-factors to repress expression of luciferase reporters driven by these sequences. Most significantly, we have found that conditional loss of Shh in limb buds lacking GATA6 prevents development of hindlimb polydactyly in these compound mutant embryos, indicating that GATA6 expression in the anterior region of the limb bud blocks hindlimb polydactyly by repressing ectopic expression of Shh.

  19. BudBurst Buddies: Introducing Young Citizen Scientists to Plants and Environmental Change

    Science.gov (United States)

    Ward, D.; Gardiner, L. S.; Henderson, S.

    2011-12-01

    As part of Project BudBurst, the BudBurst Buddies recently moved to the National Ecological Network (NEON) as part of its Education and Public Engagement efforts. The BudBurst Buddies (www.budburstbuddies.org) were created to engage elementary school age children in the science of observing plants and the timing of phenological (life cycle) events. BudBurst Buddies is a part of the Project BudBurst national citizen science initiative (www.budburst.org), which allows individuals to engage in the scientific process, contributing to a better understanding of climate change while increasing public awareness of phenology and the impacts of climate change on plants. As a first step towards engaging the next generation of citizen scientists, BudBurst Buddies provides the opportunity for children to gain experience with scientific research and increases awareness of how plants change throughout the year. Hundreds of young students have participated in the inaugural year of BudBurst Buddies. Children can participate in BudBurst Buddies on their own, with their families, or in formal or informal education settings. The program was recently highlighted by education staff at the New York Hall of Science and numerous classrooms have been implementing this resource as part of their curriculum. Each child who participates creates a journal about a plant of his or her choosing, makes observations of the plant over the growing season and submits findings online, earning an official BudBurst Buddies certificate. An online storybook for kids tells how two children, Lily and Sage, observed plants in their neighborhood and became BudBurst Buddies. This presentation will provide an overview of the BudBurst Buddies resources including a new implementation guide and will also share feedback from the first year of implementation.

  20. A Fate Map of the Murine Pancreas Buds Reveals a Multipotent Ventral Foregut Organ Progenitor

    Science.gov (United States)

    Angelo, Jesse R.; Guerrero-Zayas, Mara-Isel; Tremblay, Kimberly D.

    2012-01-01

    The definitive endoderm is the embryonic germ layer that gives rise to the budding endodermal organs including the thyroid, lung, liver and pancreas as well as the remainder of the gut tube. DiI fate mapping and whole embryo culture were used to determine the endodermal origin of the 9.5 days post coitum (dpc) dorsal and ventral pancreas buds. Our results demonstrate that the progenitors of each bud occupy distinct endodermal territories. Dorsal bud progenitors are located in the medial endoderm overlying somites 2–4 between the 2 and 11 somite stage (SS). The endoderm forming the ventral pancreas bud is found in 2 distinct regions. One territory originates from the left and right lateral endoderm caudal to the anterior intestinal portal by the 6 SS and the second domain is derived from the ventral midline of the endoderm lip (VMEL). Unlike the laterally located ventral foregut progenitors, the VMEL population harbors a multipotent progenitor that contributes to the thyroid bud, the rostral cap of the liver bud, ventral midline of the liver bud and the midline of the ventral pancreas bud in a temporally restricted manner. This data suggests that the midline of the 9.5 dpc thyroid, liver and ventral pancreas buds originates from the same progenitor population, demonstrating a developmental link between all three ventral foregut buds. Taken together, these data define the location of the dorsal and ventral pancreas progenitors in the prespecified endodermal sheet and should lead to insights into the inductive events required for pancreas specification. PMID:22815796

  1. [Effects of basic orange II on proliferation and differentiation of limb bud cells in rat embryos].

    Science.gov (United States)

    Zheng, Lixin; Feng, Jiawang; Tian, Shimin

    2015-01-01

    To explore the effects of basic orange II on proliferation and differentiation of limb bud cells. Limb bud cell were separated from SD rat embryo at 13-day gestational age, limb bud cell were exposed to basic orange II at concentrations of 0.0, 12.5, 25.0, 50.0, 100.0, 200, 0 and 400.0 mg/L in the culture medium. The effect of basic orange II on limb bud cell proliferation was detected by Cell Counting Kit-8, the effect of basic orange II on limb bud cell differentiation was assessed by Alcian Blue 8GX. With the increasing of basic orange II concentration, the proliferation and differentiation of embryo limb bud cells were poorer and poorer in vitro, and there was the dose-effect relationship. The pID50 and dLD50 of basic orange II on limb bud cells were 240.6 mg/L and 69.3 mg/L respectively. The inhibition of basic orange II on cell differentiation might exceed that on cell proliferation. Basic orange II could inhibit proliferation and differentiation of embryo limb bud cells. It might be a potential developmental toxic substance in rat embryo.

  2. Morphological and physiological aspects of the early phases of flower bud formation of apple

    NARCIS (Netherlands)

    Verheij, F.A.

    1996-01-01


    For consistent yields in apple fruit production, knowledge of the factors affecting flower bud formation is required. The aim of this study was to gain more insight in the role of endogenous factors in flower bud formation of apple. The effects of temperature, applied gibberellin (GA

  3. Thidiazuron-induced in vitro bud organogenesis of the date palm ...

    African Journals Online (AJOL)

    The objective of the present was to enhance the frequency of plant regeneration in date palm (Phoenix dactylifera L.) cv. Hillawi. ... studies revealed that adventitious buds were formed directly from epidermal cells without callus formation, and adventitious buds were developed from meristematic cells in shoot tip tissues.

  4. effects of bud loading levels and different nitrogen doses on the ...

    African Journals Online (AJOL)

    USER

    2011-09-28

    Sep 28, 2011 ... The aim of this study was to investigate the effects of several bud loading levels in winter pruning and nitrogen doses on yield and physical and chemical properties of fresh vine-leaves of grape cultivar. “Narince”. Vines trained with bilateral cordon system was pruned to yield 35000 to 53000 buds/ha (16 or.

  5. Light controls shoot meristem organogenic activity and leaf primordia growth during bud burst in Rosa sp.

    Science.gov (United States)

    Girault, Tiffanie; Bergougnoux, Veronique; Combes, Didier; Viemont, Jean-Daniel; Leduc, Nathalie

    2008-11-01

    Light controls bud burst in many plants, which subsequently affects their architecture. Nevertheless, very little is known about this photomorphogenic process. This study ascertains the effects of light on bud burst and on two of its components, i.e. growth of preformed leaves and meristem organogenesis in six cultivars from three Rosa species (R. hybrida L., R. chinensis L., R. wichurana L.). Defoliated plants were severed above the third basal bud and exposed, either to darkness or to different intensities of white light, to blue, red or to FR, at constant temperature. Bud bursting was inhibited in darkness in the six cultivars of Rosa, but not in Arabidopsis, tomato and poplar plants under the same condition. In all Rosa cultivars, bud burst, growth of preformed leaves and meristem organogenesis were triggered by blue and red lights, and extended by increasing light intensities. FR was inhibitory of bud burst. Partial shading experiments demonstrated that bud and not stem was the active site for light perception in bud burst.

  6. Loss of membranous Ep-CAM in budding colorectal carcinoma cells.

    NARCIS (Netherlands)

    Gosens, M.J.E.M.; Kempen, L.C. van; Velde, C.J. van de; Krieken, J.H.J.M. van; Nagtegaal, I.D.

    2007-01-01

    Tumor budding is a histological feature that reflects loss of adhesion of tumor cells and is associated with locoregional metastasis of colorectal carcinoma. Although nuclear localization of beta-catenin is associated with tumor budding, the molecular mechanism remains largely elusive. In this

  7. Prognostic significance of tumor budding in rectal cancer biopsies before neoadjuvant therapy.

    Science.gov (United States)

    Rogers, Ailín C; Gibbons, David; Hanly, Ann M; Hyland, John M P; O'Connell, P Ronan; Winter, Desmond C; Sheahan, Kieran

    2014-01-01

    Tumor budding is an increasingly important prognostic feature for pathologists to recognize. The aim of this study was to correlate intra-tumoral budding in pre-treatment rectal cancer biopsies with pathological response to neoadjuvant chemoradiotherapy and with long-term outcome. Data from a prospectively maintained database were acquired from patients with locally advanced rectal cancer who underwent neoadjuvant chemoradiotherapy. Pre-treatment rectal biopsies were retrospectively reviewed for evidence of intra-tumoral budding. Multivariate logistic regression was used to identify factors contributing to cancer-specific death, expressed as hazard ratios with 95% confidence intervals. Of the 185 patients with locally advanced rectal cancer, 89 patients met the eligibility criteria, of whom 18 (20%) exhibited budding in a pre-treatment tumor biopsy. Intra-tumoral budding predicted a poor pathological response to neoadjuvant chemoradiotherapy (higher ypT stage, P=0.032; lymph node involvement, P=0.018; lymphovascular invasion, P=0.004; and residual poorly differentiated tumors, P=0.005). No patient with intra-tumoral budding exhibited a tumor regression grade 1 or complete pathological response, providing a 100% specificity and positive predictive value for non-response to neoadjuvant chemoradiotherapy. Intra-tumoral budding was associated with a lower disease-free 5-year survival rate (33 vs 78%, Ptumoral budding at diagnosis of rectal cancer identifies those who will poorly respond to neoadjuvant chemoradiotherapy and those with a poor prognosis.

  8. Effect of pretreatment methods of dormant pear buds on viability after cryopreservation

    Science.gov (United States)

    This study aimed to develop alternatives for dormant bud cryopreservation by using several cryoprotectants on four pear cultivars with a view to improve the viability of the dormant buds. We used different cryoprotectants such as Honey, PVS2, PVS3, PVS4, Towill, IPBB-1 for cultivars: Talgarskaya Kra...

  9. New technique for more rapid cryopreservation of dormant vegetative tree buds

    Science.gov (United States)

    Cryopreservation of dormant buds of temperate trees in liquid nitrogen can provide a safe backup of field germplasm collections. However the process requires several months of preparation before buds can be cryopreserved. Cryopreservation at the natural moisture content (MC) would greatly accelerate...

  10. Apical bud toughness tests and tree sway movements to examine crown abrasion: preliminary results

    Science.gov (United States)

    Tyler Brannon; Wayne Clatterbuck

    2012-01-01

    Apical bud toughness differences were examined for several species to determine if crown abrasion affects shoot growth of determinate and indeterminate species during stand development. Determinate buds will set and harden after initial shoot elongation in the spring, while the indeterminate shoots form leaves from the apical meristem continuously based on the...

  11. The YPLGVG sequence of the Nipah virus matrix protein is required for budding

    Directory of Open Access Journals (Sweden)

    Yan Lianying

    2008-11-01

    Full Text Available Abstract Background Nipah virus (NiV is a recently emerged paramyxovirus capable of causing fatal disease in a broad range of mammalian hosts, including humans. Together with Hendra virus (HeV, they comprise the genus Henipavirus in the family Paramyxoviridae. Recombinant expression systems have played a crucial role in studying the cell biology of these Biosafety Level-4 restricted viruses. Henipavirus assembly and budding occurs at the plasma membrane, although the details of this process remain poorly understood. Multivesicular body (MVB proteins have been found to play a role in the budding of several enveloped viruses, including some paramyxoviruses, and the recruitment of MVB proteins by viral proteins possessing late budding domains (L-domains has become an important concept in the viral budding process. Previously we developed a system for producing NiV virus-like particles (VLPs and demonstrated that the matrix (M protein possessed an intrinsic budding ability and played a major role in assembly. Here, we have used this system to further explore the budding process by analyzing elements within the M protein that are critical for particle release. Results Using rationally targeted site-directed mutagenesis we show that a NiV M sequence YPLGVG is required for M budding and that mutation or deletion of the sequence abrogates budding ability. Replacement of the native and overlapping Ebola VP40 L-domains with the NiV sequence failed to rescue VP40 budding; however, it did induce the cellular morphology of extensive filamentous projection consistent with wild-type VP40-expressing cells. Cells expressing wild-type NiV M also displayed this morphology, which was dependent on the YPLGVG sequence, and deletion of the sequence also resulted in nuclear localization of M. Dominant-negative VPS4 proteins had no effect on NiV M budding, suggesting that unlike other viruses such as Ebola, NiV M accomplishes budding independent of MVB cellular proteins

  12. Terminal bud failure of black cottonwood (Populus trichocarpa) exposed to salt-laden winter storms.

    Science.gov (United States)

    Jonsson, Thorbergur H

    2006-07-01

    At coastal sites, trees are exposed to marine aerosols that may cause foliar necrosis and shoot dieback, which can result in deformed crowns and contorted stems. A six-year study of leaf primordia in terminal buds of black cottonwood trees (Populus trichocarpa Torr. & Gray) on Heimaey Island off the south coast of Iceland was undertaken to elucidate the physiological events associated with salt-deposition-related bud failure. Leaf and bud lengths, dry mass, water content and chloride concentrations were monitored and related to four phenological stages: (1) bud set; (2) dormancy induction; (3) dormancy release; and (4) bud break. The trees set buds in July and shed their leaves by late September. Leaf primordia generally stopped growing by September 10 +/- 22 days and attained midwinter water content in late September. Leaf growth commenced in the terminal buds by March 2 +/- 16 days, but mean dates of bud swelling and bud break were April 29 +/- 19 and May 10 +/- 12 days. In summer and until November, chloride concentrations in leaf primordia were low, but increasing. Chloride concentrations remained stable from December to February, even though the dormant trees were exposed to large amounts of marine aerosols. In February and March, three events occurred more or less simultaneously: (1) leaf extension growth commenced; (2) chloride concentration surged in the leaf primordia; and (3) the leaf primordia began to hydrate. Following dormancy release, growth and hydration of leaf primordia were negatively related to chloride concentration in the leaf primordia, with inhibition of leaf growth, tissue hydration and chloride acquisition occurring at a chloride concentration threshold estimated at 7.3 mg Cl- g(-1) tissue water. Necrosis of leaf primordia was observed above 14 mg Cl- g(-1) tissue water. Growth and hydration of leaves at bud break in mid-May was explained by a three-parameter logistic model of chloride concentration in leaf primordia at the end of March

  13. Glucosinolate composition of young shoots and flower buds of capers (Capparis species) growing wild in Turkey.

    Science.gov (United States)

    Matthäus, Bertrand; Ozcan, Musa

    2002-12-04

    The content and glucosinolate composition of young shoots and raw flower buds of Capparis spinosa var. spinosa and Capparis ovata Desf. var. canescens at three different sizes (x 13 mm) were investigated by HPLC with UV detection. Samples were harvested in August 2001 in Turkey. Twelve different glucosinolates were identified in the young shoots and buds of both species. Total content of glucosinolates ranged from 6.55 micromol/g (large buds of C. spinosa) to 45.56 micromol/g (young shoots of C. ovata). The main glucosinolate was glucocapperin, which amounted to approximately 90% of the total glucosinolates. In both species the total glucosinolate content varied in dependence on the bud size, whereas a greater variability was given for buds from C. spinosa.

  14. Cell kinetic study on the relation between irradiation hypogeusia and taste buds in rats

    Energy Technology Data Exchange (ETDEWEB)

    Kubota, Hideharu; Furumoto, Keiichi [Nippon Dental Univ., Tokyo (Japan)

    1998-12-01

    The present study was designed to elucidate the mechanism of hypogeusia caused by irradiation. X-ray treatment at 10 Gy or 20 Gy was given to the maxillofacial region including the tongue in rats, and the involvement of taste bud for hypogeusia was investigated. In addition, cytological kinetics were immunohistologically studied using bromodeoxyuridine in the taste bud and in the lingual mucosal epithelium. The following results were obtained: In the 10 Gy group, the number of taste bud become less after the exposure, but no hypogeusia was observed during the experimental period. In the 20 Gy group, any labeled taste bud was not observed on the 7th day, and all taste buds disappeared by the 10th day. In the lingual mucosal epithelium, the number of basal cells decreased to the minimum, and the body weight and total water intake decreased coincidently in the 20 Gy group, which were few in the 10 Gy group. (author)

  15. The ureteric bud epithelium: Morphogenesis and roles in metanephric kidney patterning

    Science.gov (United States)

    Nagalakshmi, Vidya K.; Yu, Jing

    2015-01-01

    The mammalian metanephric kidney is composed of two epithelial components –the collecting duct system and the nephron epithelium– that differentiate from two different tissues –the ureteric bud epithelium and the nephron progenitors, respectively– of intermediate mesoderm origin. The collecting duct system is generated through reiterative ureteric bud branching morphogenesis whereas the nephron epithelium is formed in a process termed nephrogenesis, which is initiated with the mesenchymal-epithelial transition of the nephron progenitors. Ureteric bud branching morphogenesis is regulated by nephron progenitors, and in return the ureteric bud epithelium regulates nephrogenesis. The metanephric kidney is also physiologically divided along the cortico-medullary axis into subcompartments that are enriched with specific segments of these two epithelial structures. Here we provide an overview of the major molecular and cellular processes underlying the morphogenesis and patterning of the ureteric bud epithelium and its roles in the cortical-medullary patterning of the metanephric kidney. PMID:25783232

  16. Origin of nuclear buds and micronuclei in normal and folate-deprived human lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Lindberg, Hanna K. [New Technologies and Risks, Work Environment Development, Finnish Institute of Occupational Health, Topeliuksenkatu 41aA, FI-00250 Helsinki (Finland); Wang Xu [Genome Health and Nutrigenomics Project, CSIRO Human Nutrition, Adelaide BC, SA 5000 (Australia); School of Life Sciences, Yunnan Normal University, Kunming, Yunnan 650092 (China); Jaerventaus, Hilkka [New Technologies and Risks, Work Environment Development, Finnish Institute of Occupational Health, Topeliuksenkatu 41aA, FI-00250 Helsinki (Finland); Falck, Ghita C.-M. [New Technologies and Risks, Work Environment Development, Finnish Institute of Occupational Health, Topeliuksenkatu 41aA, FI-00250 Helsinki (Finland); Norppa, Hannu [New Technologies and Risks, Work Environment Development, Finnish Institute of Occupational Health, Topeliuksenkatu 41aA, FI-00250 Helsinki (Finland)]. E-mail: hannu.norppa@ttl.fi; Fenech, Michael [Genome Health and Nutrigenomics Project, CSIRO Human Nutrition, Adelaide BC, SA 5000 (Australia)

    2007-04-01

    Micronuclei are formed from chromosomes and chromosomal fragments that lag behind in anaphase and are left outside daughter nuclei in telophase. They may also be derived from broken anaphase bridges. Nuclear buds, micronucleus-like bodies attached to the nucleus by a thin nucleoplasmic connection, have been proposed to be generated similarly to micronuclei during nuclear division or in S-phase as a stage in the extrusion of extra DNA, possibly giving rise to micronuclei. To better understand these phenomena, we have characterized the contents of 894 nuclear buds and 1392 micronuclei in normal and folate-deprived 9-day cultures of human lymphocytes using fluorescence in situ hybridization with pancentromeric and pantelomeric DNA probes. Such information has not earlier been available for human primary cells. Surprisingly, there appears to be no previous data on the occurrence of telomeres in micronuclei (or buds) of normal human cells in general. Our results suggest that nuclear buds and micronuclei have partly different mechanistic origin. Interstitial DNA without centromere or telomere label was clearly more prevalent in nuclear buds (43%) than in micronuclei (13%). DNA with only telomere label or with both centromere and telomere label was more frequent in micronuclei (62% and 22%, respectively) than in nuclear buds (44% and 10%, respectively). Folate deprivation especially increased the frequency of nuclear buds and micronuclei harboring telomeric DNA and nuclear buds harboring interstitial DNA but also buds and micronuclei with both centromeric and telomeric DNA. According to the model we propose, that micronuclei in binucleate lymphocytes primarily derive from lagging chromosomes and terminal acentric fragments during mitosis. Most nuclear buds, however, are suggested to originate from interstitial or terminal acentric fragments, possibly representing nuclear membrane entrapment of DNA that has been left in cytoplasm after nuclear division or excess DNA that

  17. Tumor budding predicts response to anti-EGFR therapies in metastatic colorectal cancer patients

    Science.gov (United States)

    Zlobec, Inti; Molinari, Francesca; Martin, Vittoria; Mazzucchelli, Luca; Saletti, Piercarlo; Trezzi, Rosangela; De Dosso, Sara; Vlajnic, Tatjana; Frattini, Milo; Lugli, Alessandro

    2010-01-01

    AIM: To investigate whether the evaluation of tumor budding can complement K-RAS analysis to improve the individualized prediction of response to anti-epidermal growth factor receptor based therapies in metastatic colorectal cancer (mCRC) patients. METHODS: Forty-three patients with mCRC treated with cetuximab or panitumumab were entered into this study. According to the Response Evaluation Criteria in Solid Tumors criteria, 30 patients had stable or progressive disease (non-responsive), while 13 patients had a partial response. Tumor buds were evaluated from whole tissue sections stained for pan-cytokeratin, evaluated in the densest region using a 40 × objective and “high-grade” tumor budding was defined as 15 buds/high-power field. RESULTS: Tumor buds and K-RAS mutation both correctly classified 68% of patients. All patients with K-RAS mutation (n = 7) or high-grade tumor budding (n = 11) were non-responsive, of which 4 patients had both features. All 13 partial responders were K-RAS wild-type with low-grade tumor budding. Combined, the predictive value of K-RAS and tumor budding was 80%. Additionally, high-grade tumor budding was significantly related to worse progression-free survival [HR (95% CI): 2.8 (1.3-6.0, P = 0.008)]. CONCLUSION: If confirmed in larger cohorts, the addition of tumor budding to K-RAS analysis may represent an effective approach for individualized patient management in the metastatic setting. PMID:20939111

  18. Prognostic Value of Tumor Budding in Early-Stage Cervical Adenocarcinomas

    Science.gov (United States)

    Satabongkoch, Nopporn; Khunamornpong, Surapan; Pongsuvareeyakul, Tip; Settakorn, Jongkolnee; Sukpan, Kornkanok; Soongkhaw, Aungsumalee; intaraphet, Suthida; Suprasert, Prapaporn; Siriaunkgul, Sumalee

    2017-06-25

    Background: Tumor budding has recently been reported as an independent adverse prognostic factor for colorectal adenocarcinomas and other types of carcinoma in the digestive tract. This study aimed to evaluate the prognostic value of tumor budding in patients with early-stage cervical adenocarcinomas and any associations with other clinical and pathological features. Methods: Histological slides of patients with early-stage (IB-IIA) usual-type endocervical adenocarcinoma who underwent radical hysterectomy and pelvic lymph node dissection, without preoperative chemotherapy, between January 2006 and December 2012 were reviewed. Tumor budding was evaluated in routinely-stained sections and defined as detached single cells or clusters of fewer than 5 cells in a tumor invasive front and was stratified based on the number of bud counts in 10-high-power fields as low (buds) and high (≥15 buds). Correlations between tumor bud count and other clinical and pathological variables including follow-up outcomes were assessed. Results: Of 129 patients, a high tumor bud count was observed in 15 (11.6%), positively associated with histologic grade 3 (ptumor size >2 cm (p=0.036). Kaplan-Meyer analysis showed a significant decrease in both disease-free survival and cancer-specific survival for patients with a high tumor bud count (p=0.027 and 0.031, respectively). On multivariate analysis, histologic grade 3 was the only independent predictor for decreased disease-free survival (p=0.004) and cancer-specific survival (p=0.003). Conclusions: A high tumor budding count based on assessment of routinely-stained sections was found to be associated with decreased disease-free and cancer-specific survival in patients with early-stage cervical adenocarcinomas. However, it was not found to be an independent prognostic predictor in this study. Creative Commons Attribution License

  19. HtrA3 stromal expression is correlated with tumor budding in stage II colorectal cancer.

    Science.gov (United States)

    Forse, Catherine L; Rahimi, Mahdi; Diamandis, Eleftherios P; Assarzadegan, Naziheh; Dawson, Heather; Grin, Andrea; Kennedy, Erin; O'Connor, Brenda; Messenger, David E; Riddell, Robert H; Kirsch, Richard; Karagiannis, George S

    2017-08-01

    Tumor budding is a well-established adverse prognostic factor in colorectal carcinoma (CRC). It may represent a form of epithelial-to-mesenchymal transition (EMT), although the underlying mechanisms remain unclear. High-temperature requirement A3 (HtrA3) is an inhibitor of the bone morphogenetic protein pathway, the suppression of which has been linked to EMT. Since HtrA3 is highly expressed in the desmoplastic stroma at the CRC invasive front, we sought to evaluate the relationship between tumor budding and HtrA3 expression in 172 stage II CRC resection specimens. All tumors were evaluated for tumor budding, with the highest budding slide selected for pan-keratin (CK) and HtrA3 immunohistochemistry. Representative areas of tumor core and invasive front, including budding and non-budding areas, were marked on CK stained slides, and then evaluated on HtrA3 stained slides. HtrA3 expression in tumor cells (tHtrA3) and peritumoral stroma (sHtrA3) was assessed for staining percentage and intensity (the product yielding a final score). Tumors with high-grade tumor budding (HGTB) showed increased expression of sHtrA3 in budding areas compared to non-budding areas at the invasive front (Ptumors compared to minimally budding tumors (Ptumor core (but not invasive front) was significantly associated with decreased 5-year overall survival on univariate analysis (P<0.05), but not multivariate analysis. HtrA3 expression in the peritumoral stroma of patients with stage II CRC is associated with HGTB and may be a novel marker of poor outcome. Copyright © 2017 Elsevier Inc. All rights reserved.

  20. Tumor budding is an independent predictor of outcome in AJCC/UICC stage II colorectal cancer.

    Science.gov (United States)

    Betge, Johannes; Kornprat, Peter; Pollheimer, Marion J; Lindtner, Richard A; Schlemmer, Andrea; Rehak, Peter; Vieth, Michael; Langner, Cord

    2012-11-01

    In colorectal cancer, the morphology of the invasive tumor margin may reflect aggressiveness of tumor growth, thus providing important prognostic information. The tumor growth pattern according to Jass and the extent of tumor budding were analyzed in patients with American Joint Committee on Cancer/Union for International Cancer Control (AJCC/UICC) stage II disease. Tumors of 120 randomly selected patients with AJCC/UICC stage II disease were retrospectively reviewed for tumor growth pattern (expanding vs. infiltrating) and the extent of tumor budding, with high-grade budding reflecting presence of 10 or more budding foci scattered at the invasive tumor margin. Progression-free and cancer-specific survivals were determined by the Kaplan-Meier method. For multivariable analysis, Cox's proportional hazards regression models were performed. The infiltrating growth pattern was significantly associated with histological subtype and lymphovascular invasion, while high-grade budding was significantly associated with tumor grade and lymphovascular invasion. High-grade budding, but not the infiltrating growth pattern, was significantly associated with outcome in univariable analysis. Cox's proportional hazards regression models proved tumor budding to be an independent predictor of disease progression (hazard ratio 3.91, 95 % confidence interval 1.3-11.77; P = 0.02) and cancer-related death (hazard ratio 5.90, 95 % confidence interval 1.62-21.51; P = 0.007). The combination of infiltrating growth pattern and high-grade budding did not have a stronger prognostic significance than tumor budding alone. Tumor budding independently predicted patient outcome in patients with AJCC/UICC stage II colorectal cancer and may therefore be used for accurate prognostication, patient counseling, and design of clinical trials by using integrated multimodal therapy.

  1. BudBurst Buddies: A New Tool for Engaging the Youngest Citizen Scientists

    Science.gov (United States)

    Gardiner, L. S.; Henderson, S.; Ward, D.

    2010-12-01

    BudBurst Buddies (www.budburstbuddies.org) introduces elementary school age children to the science of observing plants and the timing of phenological (life cycle) events. BudBurst Buddies is a new part of the Project BudBurst national citizen science initiative (www.budburst.org), which allows individuals to engage in the scientific process, contributing to a better understanding of climate change while increasing public awareness of phenology and the impacts of climate change on plants. As a first step towards engaging the next generation of citizen scientists, BudBurst Buddies provides the opportunity for children to gain experience with scientific research and increases awareness of how plants change throughout the year. Children can participate in BudBurst Buddies on their own, with their families, or in formal or informal education settings. Each child who participates creates a journal about a plant of his or her choosing, makes observations of the plant over the growing season and submits findings online, earning an official BudBurst Buddies certificate. An online storybook for kids tells how two children, Lily and Sage, observed plants in their neighborhood and became BudBurst Buddies. This presentation will provide an overview of the BudBurst Buddies newly developed resources. BudBurst Buddies is a part of Project BudBurst, a national citizen science program coordinated by the National Ecological Observatory Network (NEON) and the Chicago Botanic Garden. Funding for this resource was provided by NEON, NSF, NASA, and the National Geographic Education Foundation.

  2. Correlation between tumor budding and post-resection prognosis in patients with invasive squamous cell carcinoma of the thoracic esophagus.

    Science.gov (United States)

    Nakanishi, Yoshitsugu; Ohara, Masanori; Doumen, Hiromitsu; Kimura, Noriko; Ishidate, Takuzo; Kondo, Satoshi

    2011-02-01

    Tumor budding is defined as an isolated single cancer cell or a cluster of cancer cells composed of fewer than five cells at the invasive front of a tumor. The aim of the present study was to identify correlations between tumor budding and clinicopathological factors and their impact on postoperative prognosis in invasive squamous cell carcinoma of the thoracic esophagus. The subjects were 82 patients who underwent curative resection. The number of tumors showing budding was counted immunohistochemically based on positivity for cytokeratin (AE1/AE3) to distinguish cancer cells from inflammatory cells, and the patients were divided into low-grade and high-grade budding groups. High-grade budding was defined as ≥5 budding foci per field under a 20× objective lens. One group of 36 patients (43.9%) and a second group of 46 patients (56.1%) were classified into the low-grade and high-grade budding groups, respectively. Patients in the high-grade budding group had significantly larger tumor size, deeper depth of invasion, more frequent lymphatic invasion and venous invasion, as well as more lymph node metastases, than those in the low-grade budding group. Disease-free and overall survivals were worse in the high-grade budding group than in the low-grade budding group (Ptumor recurrence more than 5 years postoperatively belonged to the low-grade budding group. Tumor budding appears to be an indicator of tumor growth and invasiveness. Patients with low-grade budding are likely to have a better prognosis than those with high-grade budding, but the tumor may recur in the late phase postoperatively in patients with low-grade budding.

  3. Guillaume Budé, l’humaniste et le prince

    Directory of Open Access Journals (Sweden)

    Sylvie Le Clech-Charton

    2009-09-01

    Full Text Available Grande figure de la Renaissance des lettres et des arts en France, tout à la fois écrivain, traducteur, ambassadeur, créateur du dépôt légal et fondateur du Collège de France, maître de la librairie du roi à Fontainebleau, Guillaume Budé (1468-1540 est essentiellement connu pour le rôle de conseiller politique et culturel qu’il joua auprès de François Ier, dont il fut le secrétaire. Il a été surtout étudié du point de vue de sa production littéraire savante, mais non sous l’angle de son mili...

  4. Identification of SUMO conjugation sites in the budding yeast proteome

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    Miguel Esteras

    2017-10-01

    Full Text Available Post-translational modification by the small ubiquitin-like modifier (SUMO is an important mechanism regulating protein function. Identification of SUMO conjugation sites on substrates is a challenging task. Here we employed a proteomic method to map SUMO acceptor lysines in budding yeast proteins. We report the identification of 257 lysine residues where SUMO is potentially attached. Amongst the hits, we identified already known SUMO substrates and sites, confirming the success of the approach. In addition, we tested several of the novel substrates using SUMO immunoprecipitation analysis and confirmed that the SUMO acceptor lysines identified in these proteins are indeed bona fide SUMOylation sites. We believe that the collection of SUMO sites presented here is an important resource for future functional studies of SUMOylation in yeast.

  5. Genetic bypass of essential RNA repair enzymes in budding yeast.

    Science.gov (United States)

    Cherry, Patrick D; White, Laura K; York, Kerri; Hesselberth, Jay R

    2017-12-06

    RNA repair enzymes catalyze rejoining of an RNA molecule after cleavage of phosphodiester linkages. RNA repair in budding yeast is catalyzed by two separate enzymes that process tRNA exons during their splicing and HAC1 mRNA exons during activation of the unfolded protein response. The RNA ligase Trl1 joins 2',3'-cyclic phosphate and 5'-hydroxyl RNA fragments, creating a new phosphodiester linkage with a 2'-phosphate at the junction. The 2'-phosphate is subsequently removed by the 2'-phosphotransferase Tpt1, which catalyzes phosphate transfer to NAD+, producing nicotinamide and a unique ADP ribose metabolite. We bypassed the essential functions of TRL1 and TPT1 in budding yeast by expressing "pre-spliced," intronless versions of the ten normally intron-containing tRNAs, indicating this repair pathway does not have additional essential functions. Consistent with previous studies, expression of intronless tRNAs failed to rescue the growth of cells with deletions in components of the SEN complex, implying an additional essential role for the splicing endonuclease. The trl1∆ and tpt1∆ mutants accumulate tRNA and HAC1 splicing intermediates indicative of specific RNA repair defects and are hypersensitive to drugs that inhibit translation. As expected, failure to induce the unfolded protein response in trl1∆ cells grown with tunicamycin is lethal owing to their inability to ligate HAC1 after its cleavage by Ire1. In contrast, tpt1∆ mutants grow in the presence of tunicamycin despite reduced accumulation of spliced HAC1, suggesting that ligated but 2'-phosphorylated mRNA is decoded by the ribosome. Finally, we optimized a PCR-based method to detect RNA 2'-phosphate modifications and show that they are present on ligated HAC1 mRNA. These RNA repair mutants enable new studies of the role of RNA repair in cellular physiology. Published by Cold Spring Harbor Laboratory Press for the RNA Society.

  6. Sonic hedgehog from both nerves and epithelium is a key trophic factor for taste bud maintenance.

    Science.gov (United States)

    Castillo-Azofeifa, David; Losacco, Justin T; Salcedo, Ernesto; Golden, Erin J; Finger, Thomas E; Barlow, Linda A

    2017-09-01

    The integrity of taste buds is intimately dependent on an intact gustatory innervation, yet the molecular nature of this dependency is unknown. Here, we show that differentiation of new taste bud cells, but not progenitor proliferation, is interrupted in mice treated with a hedgehog (Hh) pathway inhibitor (HPI), and that gustatory nerves are a source of sonic hedgehog (Shh) for taste bud renewal. Additionally, epithelial taste precursor cells express Shh transiently, and provide a local supply of Hh ligand that supports taste cell renewal. Taste buds are minimally affected when Shh is lost from either tissue source. However, when both the epithelial and neural supply of Shh are removed, taste buds largely disappear. We conclude Shh supplied by taste nerves and local taste epithelium act in concert to support continued taste bud differentiation. However, although neurally derived Shh is in part responsible for the dependence of taste cell renewal on gustatory innervation, neurotrophic support of taste buds likely involves a complex set of factors. © 2017. Published by The Company of Biologists Ltd.

  7. Evolutionary origins of taste buds: phylogenetic analysis of purinergic neurotransmission in epithelial chemosensors

    Science.gov (United States)

    Kirino, Masato; Parnes, Jason; Hansen, Anne; Kiyohara, Sadao; Finger, Thomas E.

    2013-01-01

    Taste buds are gustatory endorgans which use an uncommon purinergic signalling system to transmit information to afferent gustatory nerve fibres. In mammals, ATP is a crucial neurotransmitter released by the taste cells to activate the afferent nerve fibres. Taste buds in mammals display a characteristic, highly specific ecto-ATPase (NTPDase2) activity, suggesting a role in inactivation of the neurotransmitter. The purpose of this study was to test whether the presence of markers of purinergic signalling characterize taste buds in anamniote vertebrates and to test whether similar purinergic systems are employed by other exteroceptive chemosensory systems. The species examined include several teleosts, elasmobranchs, lampreys and hagfish, the last of which lacks vertebrate-type taste buds. For comparison, Schreiner organs of hagfish and solitary chemosensory cells (SCCs) of teleosts, both of which are epidermal chemosensory end organs, were also examined because they might be evolutionarily related to taste buds. Ecto-ATPase activity was evident in elongate cells in all fish taste buds, including teleosts, elasmobranchs and lampreys. Neither SCCs nor Schreiner organs show specific ecto-ATPase activity, suggesting that purinergic signalling is not crucial in those systems as it is for taste buds. These findings suggest that the taste system did not originate from SCCs but arose independently in early vertebrates. PMID:23466675

  8. Automated quantification of budding Saccharomyces cerevisiae using a novel image cytometry method.

    Science.gov (United States)

    Laverty, Daniel J; Kury, Alexandria L; Kuksin, Dmitry; Pirani, Alnoor; Flanagan, Kevin; Chan, Leo Li-Ying

    2013-06-01

    The measurements of concentration, viability, and budding percentages of Saccharomyces cerevisiae are performed on a routine basis in the brewing and biofuel industries. Generation of these parameters is of great importance in a manufacturing setting, where they can aid in the estimation of product quality, quantity, and fermentation time of the manufacturing process. Specifically, budding percentages can be used to estimate the reproduction rate of yeast populations, which directly correlates with metabolism of polysaccharides and bioethanol production, and can be monitored to maximize production of bioethanol during fermentation. The traditional method involves manual counting using a hemacytometer, but this is time-consuming and prone to human error. In this study, we developed a novel automated method for the quantification of yeast budding percentages using Cellometer image cytometry. The automated method utilizes a dual-fluorescent nucleic acid dye to specifically stain live cells for imaging analysis of unique morphological characteristics of budding yeast. In addition, cell cycle analysis is performed as an alternative method for budding analysis. We were able to show comparable yeast budding percentages between manual and automated counting, as well as cell cycle analysis. The automated image cytometry method is used to analyze and characterize corn mash samples directly from fermenters during standard fermentation. Since concentration, viability, and budding percentages can be obtained simultaneously, the automated method can be integrated into the fermentation quality assurance protocol, which may improve the quality and efficiency of beer and bioethanol production processes.

  9. Cell cycle genes are activated earlier than respiratory genes during release of grapevine buds from endodormancy.

    Science.gov (United States)

    Noriega, Ximena; Pérez, Francisco J

    2017-10-03

    Single-bud cuttings of Vitis vinifera L exposed to forced growing conditions were used to investigate the involvement of phytohormones, abscisic acid (ABA), auxin (Aux) and cytokinin (CK) in the release of buds from the ED and in bud-sprouting. This artificial system imitates and hastens the natural sprouting that occurs in spring. Temporal expression analysis of genes related to phytohormones synthesis, showed an early drop in the expression of ABA biosynthesis gene that preceded an increase in Aux and CK biosynthesis genes. Bud-break is headed by the activation of all structures of the latent bud, especially the differentiation of the inflorescence and the development of the early stages of floral organs. Therefore, resumption of cell division and increases in respiration are essential for the activation of the bud. Temporal expression analysis of the cell cycle and respiration genes indicate that an increase in cell division go before the increase in respiration. These results, together with results indicating that the cell cycle genes are upregulated by Aux and CK, suggest that the events before the bud-break, start with a reduction in ABA content, followed by an increase in the content of Aux and CK, which activates the machinery of the cell cycle, which eventually would cause an increase in respiration.

  10. Sugars are under light control during bud burst in Rosa sp.

    Science.gov (United States)

    Girault, Tiffanie; Abidi, Farouk; Sigogne, Monique; Pelleschi-Travier, Sandrine; Boumaza, Rachid; Sakr, Soulaiman; Leduc, Nathalie

    2010-08-01

    Bud burst in certain species is conditioned by the luminous environment. With roses, the requirement for light is absolute, and darkness totally inhibits bud burst. Few studies have looked into understanding the action of light on the physiological bud burst processes. Here, we show the impact of light on certain components of glucidic metabolism during bud burst. Measurements were taken on decapitated plants of Rosa hybrida L. 'Radrazz' exposed either to darkness, white, blue or R light. Results show that a mobilization of bud and the carrying stem sucrose reserves only takes place in light and accompanies the bud burst. Furthermore, the activity of the RhVI vacuolar acid invertase which contributes to the breakdown of sucrose in the buds, as well as the transcription of the RhVI gene, is reduced in darkness, although it is strongly stimulated by light. The same analysis concerning the RhNAD-SDH gene, coding an NAD-dependent sorbitol dehydrogenase, shows, on the contrary, a strong induction of its transcription in darkness that could reflect the use of survival mechanisms in this condition.

  11. Prognostic significance of tumor budding and single cell invasion in gastric adenocarcinoma.

    Science.gov (United States)

    Che, Keying; Zhao, Yang; Qu, Xiao; Pang, Zhaofei; Ni, Yang; Zhang, Tiehong; Du, Jiajun; Shen, Hongchang

    2017-01-01

    Gastric carcinoma (GC) is a highly aggressive cancer and one of the leading causes of cancer-related deaths worldwide. Histopathological evaluation pertaining to invasiveness is likely to provide additional information in relation to patient outcome. In this study, we aimed to evaluate the prognostic significance of tumor budding and single cell invasion in gastric adenocarcinoma. Hematoxylin and eosin-stained slides generated from 296 gastric adenocarcinoma patients with full clinical and pathological and follow-up information were systematically reviewed. The patients were grouped on the basis of tumor budding, single cell invasion, large cell invasion, mitotic count, and fibrosis. The association between histopathological parameters, different classification systems, and overall survival (OS) was statistically analyzed. Among the 296 cases that were analyzed, high-grade tumor budding was observed in 49.0% (145) of them. Single cell invasion and large cell invasion were observed in 62.8% (186) and 16.9% (50) of the cases, respectively. Following univariate analysis, patients with high-grade tumor budding had shorter OS than those with low-grade tumor budding (hazard ratio [HR]: 2.260, Ptumor budding and single cell invasion were observed to be independent risk factors for gastric adenocarcinoma (PTumor budding and single cell invasion in gastric adenocarcinoma are associated with an unfavorable prognosis.

  12. Thyroid bud morphogenesis requires CDC42- and SHROOM3-dependent apical constriction.

    Science.gov (United States)

    Loebel, David A F; Plageman, Timothy F; Tang, Theresa L; Jones, Vanessa J; Muccioli, Maria; Tam, Patrick P L

    2016-01-15

    Early development of the gut endoderm and its subsequent remodeling for the formation of organ buds are accompanied by changes to epithelial cell shape and polarity. Members of the Rho-related family of small GTPases and their interacting proteins play multiple roles in regulating epithelial morphogenesis. In this study we examined the role of Cdc42 in foregut development and organ bud formation. Ablation of Cdc42 in post-gastrulation mouse embryos resulted in a loss of apical-basal cell polarity and columnar epithelial morphology in the ventral pharyngeal endoderm, in conjunction with a loss of apical localization of the known CDC42 effector protein PARD6B. Cell viability but not proliferation in the foregut endoderm was impaired. Outgrowth of the liver, lung and thyroid buds was severely curtailed in Cdc42-deficient embryos. In particular, the thyroid bud epithelium did not display the apical constriction that normally occurs concurrently with the outgrowth of the bud into the underlying mesenchyme. SHROOM3, a protein that interacts with Rho GTPases and promotes apical constriction, was strongly expressed in the thyroid bud and its sub-cellular localization was disrupted in Cdc42-deficient embryos. In Shroom3 gene trap mutant embryos, the thyroid bud epithelium showed no apical constriction, while the bud continued to grow and protruded into the foregut lumen. Our findings indicate that Cdc42 is required for epithelial polarity and organization in the endoderm and for apical constriction in the thyroid bud. It is possible that the function of CDC42 is partly mediated by SHROOM3. © 2016. Published by The Company of Biologists Ltd.

  13. Thyroid bud morphogenesis requires CDC42- and SHROOM3-dependent apical constriction

    Directory of Open Access Journals (Sweden)

    David A. F. Loebel

    2016-02-01

    Full Text Available Early development of the gut endoderm and its subsequent remodeling for the formation of organ buds are accompanied by changes to epithelial cell shape and polarity. Members of the Rho-related family of small GTPases and their interacting proteins play multiple roles in regulating epithelial morphogenesis. In this study we examined the role of Cdc42 in foregut development and organ bud formation. Ablation of Cdc42 in post-gastrulation mouse embryos resulted in a loss of apical-basal cell polarity and columnar epithelial morphology in the ventral pharyngeal endoderm, in conjunction with a loss of apical localization of the known CDC42 effector protein PARD6B. Cell viability but not proliferation in the foregut endoderm was impaired. Outgrowth of the liver, lung and thyroid buds was severely curtailed in Cdc42-deficient embryos. In particular, the thyroid bud epithelium did not display the apical constriction that normally occurs concurrently with the outgrowth of the bud into the underlying mesenchyme. SHROOM3, a protein that interacts with Rho GTPases and promotes apical constriction, was strongly expressed in the thyroid bud and its sub-cellular localization was disrupted in Cdc42-deficient embryos. In Shroom3 gene trap mutant embryos, the thyroid bud epithelium showed no apical constriction, while the bud continued to grow and protruded into the foregut lumen. Our findings indicate that Cdc42 is required for epithelial polarity and organization in the endoderm and for apical constriction in the thyroid bud. It is possible that the function of CDC42 is partly mediated by SHROOM3.

  14. Cytokinins Are Initial Targets of Light in the Control of Bud Outgrowth.

    Science.gov (United States)

    Roman, Hanaé; Girault, Tiffanie; Barbier, François; Péron, Thomas; Brouard, Nathalie; Pěnčík, Aleš; Novák, Ondřej; Vian, Alain; Sakr, Soulaiman; Lothier, Jérémy; Le Gourrierec, José; Leduc, Nathalie

    2016-09-01

    Bud outgrowth is controlled by environmental and endogenous factors. Through the use of the photosynthesis inhibitor norflurazon and of masking experiments, evidence is given here that light acts mainly as a morphogenic signal in the triggering of bud outgrowth and that initial steps in the light signaling pathway involve cytokinins (CKs). Indeed, in rose (Rosa hybrida), inhibition of bud outgrowth by darkness is suppressed solely by the application of CKs. In contrast, application of sugars has a limited effect. Exposure of plants to white light (WL) induces a rapid (after 3-6 h of WL exposure) up-regulation of CK synthesis (RhIPT3 and RhIPT5), of CK activation (RhLOG8), and of CK putative transporter RhPUP5 genes and to the repression of the CK degradation RhCKX1 gene in the node. This leads to the accumulation of CKs in the node within 6 h and in the bud at 24 h and to the triggering of bud outgrowth. Molecular analysis of genes involved in major mechanisms of bud outgrowth (strigolactone signaling [RwMAX2], metabolism and transport of auxin [RhPIN1, RhYUC1, and RhTAR1], regulation of sugar sink strength [RhVI, RhSUSY, RhSUC2, and RhSWEET10], and cell division and expansion [RhEXP and RhPCNA]) reveal that, when supplied in darkness, CKs up-regulate their expression as rapidly and as intensely as WL Additionally, up-regulation of CKs by WL promotes xylem flux toward the bud, as evidenced by Methylene Blue accumulation in the bud after CK treatment in the dark. Altogether, these results suggest that CKs are initial components of the light signaling pathway that controls the initiation of bud outgrowth. © 2016 American Society of Plant Biologists. All rights reserved.

  15. [Clinical significance of tumor budding detection in stage II colon cancer].

    Science.gov (United States)

    Liu, Shao-jie; Yang, Xiao-hong; Ren, Jing-qing; Zhu, Xuan-jin

    2013-08-01

    To investigate the association of tumor budding with recurrence and survival of patients with stage II colon cancer, in order to identify patients with high-risk recurrence who may benefit from adjuvant therapy. Clinical data of 112 stage II colon cancer patients in Guangzhou Red Cross Hospital between 1998 and 2007 were analyzed retrospectively. The degree of tumor budding was assessed by two observers and classified according to the number of tumor buds in the area with the greatest budding intensity on HE stain slides, as high-grade budding (≥10, n=30) and low-grade budding (≤9, n=82). Progression-free and cancer-specific survival were analyzed using the Kaplan-Meier method and Cox regression. All the patients were followed up and the median follow-up was 78 months. The 5-year progression-free survival rates for patients with high-grade and low-grade budding were 65.3% and 90.7% respectively (P=0.008). The 5-year cancer-specific survival rates were 72.1% and 93.8% respectively (P=0.001). Cox regression analysis demonstrated tumor budding was an independent predictor of disease progression (RR=4.572, 95%CI:2.218-11.746, P=0.002) and cancer-related death (RR=4.116, 95%CI:1.657-10.384, P=0.012). Tumor budding is a strong prognostic index for adverse outcome in stage II colon cancer patients,which may serve as a prognostic marker to identify patients with high risk of recurrence who may benefit from adjuvant therapy.

  16. Limb patterning genes and heterochronic development of the emu wing bud

    Directory of Open Access Journals (Sweden)

    Craig A. Smith

    2016-12-01

    Full Text Available Abstract Background The forelimb of the flightless emu is a vestigial structure, with greatly reduced wing elements and digit loss. To explore the molecular and cellular mechanisms associated with the evolution of vestigial wings and loss of flight in the emu, key limb patterning genes were examined in developing embryos. Methods Limb development was compared in emu versus chicken embryos. Immunostaining for cell proliferation markers was used to analyze growth of the emu forelimb and hindlimb buds. Expression patterns of limb patterning genes were studied, using whole-mount in situ hybridization (for mRNA localization and RNA-seq (for mRNA expression levels. Results The forelimb of the emu embryo showed heterochronic development compared to that in the chicken, with the forelimb bud being retarded in its development. Early outgrowth of the emu forelimb bud is characterized by a lower level of cell proliferation compared the hindlimb bud, as assessed by PH3 immunostaining. In contrast, there were no obvious differences in apoptosis in forelimb versus hindlimb buds (cleaved caspase 3 staining. Most key patterning genes were expressed in emu forelimb buds similarly to that observed in the chicken, but with smaller expression domains. However, expression of Sonic Hedgehog (Shh mRNA, which is central to anterior–posterior axis development, was delayed in the emu forelimb bud relative to other patterning genes. Regulators of Shh expression, Gli3 and HoxD13, also showed altered expression levels in the emu forelimb bud. Conclusions These data reveal heterochronic but otherwise normal expression of most patterning genes in the emu vestigial forelimb. Delayed Shh expression may be related to the small and vestigial structure of the emu forelimb bud. However, the genetic mechanism driving retarded emu wing development is likely to rest within the forelimb field of the lateral plate mesoderm, predating the expression of patterning genes.

  17. Morphogenetic and developmental functions of the Aspergillus nidulans homologues of the yeast bud site selection proteins Bud4 and Axl2.

    Science.gov (United States)

    Si, Haoyu; Rittenour, William R; Xu, Kaimei; Nicksarlian, Mark; Calvo, Ana M; Harris, Steven D

    2012-07-01

    The yeast bud site selection system represents a paradigm for understanding how fungal cells regulate the formation of a polarity axis. In Saccharomyces cerevisiae, Bud4 and Axl2 are components of the axial bud site marker. To address the possibility that these proteins regulate cellular morphogenesis in filamentous fungi, we have characterized homologues of Bud4 and Axl2 in Aspergillus nidulans. Our results show that Bud4 is involved in septum formation in both hyphae and developing conidiophores. Whereas Axl2 appears to have no obvious role in hyphal growth, it is required for the regulation of phialide morphogenesis during conidiation. In particular, Axl2 localizes to the phialide-spore junction, where it appears to promote the recruitment of septins. Furthermore, the developmental regulators BrlA and AbaA control the expression of Axl2. Additional studies indicate that Axl2 is also involved in the regulation of sexual development, not only in A. nidulans, but also in the phylogenetically unrelated fungus Fusarium graminearum. Our results suggest that Axl2 plays a key role in phialide morphogenesis and/or function during conidiation in the aspergilli. © 2012 Blackwell Publishing Ltd.

  18. Phenotypic plasticity, QTL mapping and genomic characterization of bud set in black poplar

    Directory of Open Access Journals (Sweden)

    Fabbrini Francesco

    2012-04-01

    Full Text Available Abstract Background The genetic control of important adaptive traits, such as bud set, is still poorly understood in most forest trees species. Poplar is an ideal model tree to study bud set because of its indeterminate shoot growth. Thus, a full-sib family derived from an intraspecific cross of P. nigra with 162 clonally replicated progeny was used to assess the phenotypic plasticity and genetic variation of bud set in two sites of contrasting environmental conditions. Results Six crucial phenological stages of bud set were scored. Night length appeared to be the most important signal triggering the onset of growth cessation. Nevertheless, the effect of other environmental factors, such as temperature, increased during the process. Moreover, a considerable role of genotype × environment (G × E interaction was found in all phenological stages with the lowest temperature appearing to influence the sensitivity of the most plastic genotypes. Descriptors of growth cessation and bud onset explained the largest part of phenotypic variation of the entire process. Quantitative trait loci (QTL for these traits were detected. For the four selected traits (the onset of growth cessation (date2.5, the transition from shoot to bud (date1.5, the duration of bud formation (subproc1 and bud maturation (subproc2 eight and sixteen QTL were mapped on the maternal and paternal map, respectively. The identified QTL, each one characterized by small or modest effect, highlighted the complex nature of traits involved in bud set process. Comparison between map location of QTL and P. trichocarpa genome sequence allowed the identification of 13 gene models, 67 bud set-related expressional and six functional candidate genes (CGs. These CGs are functionally related to relevant biological processes, environmental sensing, signaling, and cell growth and development. Some strong QTL had no obvious CGs, and hold great promise to identify unknown genes that affect bud set

  19. Nerve-independent and ectopically additional induction of taste buds in organ culture of fetal tongues.

    Science.gov (United States)

    Honda, Kotaro; Tomooka, Yasuhiro

    2016-10-01

    An improved organ culture system allowed to observe morphogenesis of mouse lingual papillae and taste buds relatively for longer period, in which fetal tongues were analyzed for 6 d. Taste cells were defined as eosinophobic epithelial cells expressing CK8 and Sox2 within lingual epithelium. Addition of glycogen synthase kinase 3 beta inhibitor CHIR99021 induced many taste cells and buds in non-gustatory and gustatory stratified lingual epithelium. The present study clearly demonstrated induction of taste cells and buds ectopically and without innervation.

  20. Tumor budding is an independent prognostic factor for prediction of lymph node metastasis in oral squamous cell carcinoma.

    Science.gov (United States)

    Angadi, Punnya V; Patil, Prakash V; Hallikeri, Kaveri; Mallapur, M D; Hallikerimath, Seema; Kale, Alka D

    2015-04-01

    Despite the enormous advances in diagnostic and management modalities of oral squamous cell carcinoma (OSCC), the mortality rates have remained stagnant with a 5-year survival rate of tumor budding has been associated with aggressive behavior and is correlated with lymph node metastasis, recurrence, distant metastasis, and decreased survival in several cancers. However, the prognostic significance of this apparently simple to evaluate parameter is sparse in OSCC. A total of 75 cases of surgically excised OSCC were analyzed for tumor budding along with other clinicopathologic parameters. Tumor budding was graded as high and low intensity based on presence and absence of ≥10 or budding foci in hematoxylin and eosin-stained sections. An association between the clinicopathological parameters, lymph node metastases with the budding index was examined using univariate and multivariate analyses. Tumor budding was evident in 89% of cases with around 45.3% of the cases demonstrated high-intensity budding. High-intensity tumor budding was significantly associated with lymph node metastasis and depth of invasion. Multivariate analysis demonstrated that tumor budding and depth of invasion were significant independent predictors for lymph node metastasis. Tumor budding is frequently encountered histologic marker in OSCC. High-intensity tumor budding is a strong independent prognostic factor for prediction of lymph node metastasis. © The Author(s) 2015.

  1. Proposal for a 10-high-power-fields scoring method for the assessment of tumor budding in colorectal cancer.

    Science.gov (United States)

    Karamitopoulou, Eva; Zlobec, Inti; Kölzer, Viktor; Kondi-Pafiti, Agathi; Patsouris, Efstratios S; Gennatas, Konstantin; Lugli, Alessandro

    2013-02-01

    Although tumor budding is linked to adverse prognosis in colorectal cancer, it remains largely unreported in daily diagnostic work due to the absence of a standardized scoring method. Our aim was to assess the inter-observer agreement of a novel 10-high-power-fields method for assessment of tumor budding at the invasive front and to confirm the prognostic value of tumor budding in our setting of colorectal cancers. Whole tissue sections of 215 colorectal cancers with full clinico-pathological and follow-up information were stained with cytokeratin AE1/AE3 antibody. Presence of buds was scored across 10-high-power fields at the invasive front by two pathologists and two additional observers were asked to score 50 cases of tumor budding randomly selected from the larger cohort. The measurements were correlated to the patient and tumor characteristics. Inter-observer agreement and correlation between observers' scores were excellent (Ptumor budding and the remaining 70% of the patients were entered into the analysis. High-grade budding was defined as an average of ≥10 buds across 10-high-power fields. High-grade budding was associated with a higher tumor grade (Ptumor border configuration (Ptumor budding has independent prognostic value and shows excellent inter-observer agreement. Like the BRE and Gleason scores in breast and prostate cancers, respectively, tumor budding could be a basis for a prognostic score in colorectal cancer.

  2. Voltage-gated sodium channels in taste bud cells

    Directory of Open Access Journals (Sweden)

    Williams Mark E

    2009-03-01

    Full Text Available Abstract Background Taste bud cells transmit information regarding the contents of food from taste receptors embedded in apical microvilli to gustatory nerve fibers innervating basolateral membranes. In particular, taste cells depolarize, activate voltage-gated sodium channels, and fire action potentials in response to tastants. Initial cell depolarization is attributable to sodium influx through TRPM5 in sweet, bitter, and umami cells and an undetermined cation influx through an ion channel in sour cells expressing PKD2L1, a candidate sour taste receptor. The molecular identity of the voltage-gated sodium channels that sense depolarizing signals and subsequently initiate action potentials coding taste information to gustatory nerve fibers is unknown. Results We describe the molecular and histological expression profiles of cation channels involved in electrical signal transmission from apical to basolateral membrane domains. TRPM5 was positioned immediately beneath tight junctions to receive calcium signals originating from sweet, bitter, and umami receptor activation, while PKD2L1 was positioned at the taste pore. Using mouse taste bud and lingual epithelial cells collected by laser capture microdissection, SCN2A, SCN3A, and SCN9A voltage-gated sodium channel transcripts were expressed in taste tissue. SCN2A, SCN3A, and SCN9A were expressed beneath tight junctions in subsets of taste cells. SCN3A and SCN9A were expressed in TRPM5 cells, while SCN2A was expressed in TRPM5 and PKD2L1 cells. HCN4, a gene previously implicated in sour taste, was expressed in PKD2L1 cells and localized to cell processes beneath the taste pore. Conclusion SCN2A, SCN3A and SCN9A voltage-gated sodium channels are positioned to sense initial depolarizing signals stemming from taste receptor activation and initiate taste cell action potentials. SCN2A, SCN3A and SCN9A gene products likely account for the tetrodotoxin-sensitive sodium currents in taste receptor cells.

  3. Tumor Budding in Colorectal Carcinoma: Confirmation of Prognostic Significance and Histologic Cutoff in a Population-based Cohort.

    Science.gov (United States)

    Graham, Rondell P; Vierkant, Robert A; Tillmans, Lori S; Wang, Alice H; Laird, Peter W; Weisenberger, Daniel J; Lynch, Charles F; French, Amy J; Slager, Susan L; Raissian, Yassaman; Garcia, Joaquin J; Kerr, Sarah E; Lee, Hee Eun; Thibodeau, Stephen N; Cerhan, James R; Limburg, Paul J; Smyrk, Thomas C

    2015-10-01

    Tumor budding in colorectal carcinoma has been associated with poor outcome in multiple studies, but the absence of an established histologic cutoff for "high" tumor budding, heterogeneity in study populations, and varying methods for assessing tumor budding have hindered widespread incorporation of this parameter in clinical reports. We used an established scoring system in a population-based cohort to determine a histologic cutoff for "high" tumor budding and confirm its prognostic significance. We retrieved hematoxylin and eosin-stained sections from 553 incident colorectal carcinoma cases. Each case was previously characterized for select molecular alterations and survival data. Interobserver agreement was assessed between 2 gastrointestinal pathologists and a group of 4 general surgical pathologists. High budding (≥ 10 tumor buds in a ×20 objective field) was present in 32% of cases, low budding in 46%, and no budding in 22%. High tumor budding was associated with advanced pathologic stage (P 2 times risk of cancer-specific death (hazard ratio = 2.57 [1.27, 5.19]). After multivariate adjustment, by penalized smoothing splines, we found increasing tumor bud counts from 5 upward to be associated with an increasingly shortened cancer-specific survival. By this method, a tumor bud count of 10 corresponded to approximately 2.5 times risk of cancer-specific death. The interobserver agreement was good with weighted κ of 0.70 for 2 gastrointestinal pathologists over 121 random cases and 0.72 between all 6 pathologists for 20 random cases. Using an established method to assess budding on routine histologic stains, we have shown that a cutoff of 10 for high tumor budding is independently associated with a significantly worse prognosis. The reproducibility data provide support for the routine widespread implementation of tumor budding in clinical reports.

  4. Arf3p GTPase is a key regulator of Bud2p activation for invasive growth in Saccharomyces cerevisiae.

    Science.gov (United States)

    Hsu, Jia-Wei; Lee, Fang-Jen S

    2013-08-01

    The regulation and signaling pathways involved in the invasive growth of yeast have been studied extensively because of their general applicability to fungal pathogenesis. Bud2p, which functions as a GTPase-activating protein (GAP) for Bud1p/Rsr1p, is required for appropriate budding patterns and filamentous growth. The regulatory mechanisms leading to Bud2p activation, however, are poorly understood. In this study, we report that ADP-ribosylation factor 3p (Arf3p) acts as a regulator of Bud2p activation during invasive growth. Arf3p binds directly to the N-terminal region of Bud2p and promotes its GAP activity both in vitro and in vivo. Genetic analysis shows that deletion of BUD1 suppresses the defect of invasive growth in arf3Δ or bud2Δ cells. Lack of Arf3p, like that of Bud2p, causes the intracellular accumulation of Bud1p-GTP. The Arf3p-Bud2p interaction is important for invasive growth and facilitates the Bud2p-Bud1p association in vivo. Finally, we show that under glucose depletion-induced invasion conditions in yeast, more Arf3p is activated to the GTP-bound state, and the activation is independent of Arf3p guanine nucleotide-exchange factor Yel1p. Thus we demonstrate that a novel spatial activation of Arf3p plays a role in regulating Bud2p activation during glucose depletion-induced invasive growth.

  5. Intratumoral budding as a potential parameter of tumor progression in mismatch repair-proficient and mismatch repair-deficient colorectal cancer patients.

    Science.gov (United States)

    Lugli, Alessandro; Vlajnic, Tatjana; Giger, Olivier; Karamitopoulou, Eva; Patsouris, Efstratios S; Peros, George; Terracciano, Luigi M; Zlobec, Inti

    2011-12-01

    In colorectal cancer, tumor budding at the invasive front (peritumoral budding) is an established prognostic parameter and decreased in mismatch repair-deficient tumors. In contrast, the clinical relevance of tumor budding within the tumor center (intratumoral budding) is not yet known. The aim of the study was to determine the correlation of intratumoral budding with peritumoral budding and mismatch repair status and the prognostic impact of intratumoral budding using 2 independent patient cohorts. Following pancytokeratin staining of whole-tissue sections and multiple-punch tissue microarrays, 2 independent cohorts (group 1: n = 289; group 2: n = 222) with known mismatch repair status were investigated for intratumoral budding and peritumoral budding. In group 1, intratumoral budding was strongly correlated to peritumoral budding (r = 0.64; P budding was associated with right-sided location (P = .024), advanced T stage (P = .001) and N stage pN (P tumor margin (P = .003), and shorter survival time (P = .014). In mismatch repair-deficient cancers, high intratumoral budding was linked to higher tumor grade (P = .004), vascular invasion (P = .009), infiltrating tumor margin (P = .005), and more unfavorable survival time (P = .09). These associations were confirmed in group 2. High-grade intratumoral budding was a poor prognostic factor in univariate (P budding is an independent prognostic factor, supporting the future investigation of intratumoral budding in larger series of both preoperative and postoperative rectal and colon cancer specimens. Copyright © 2011 Elsevier Inc. All rights reserved.

  6. Transcriptome analysis of female and male flower buds of Idesia polycarpa Maxim. var. vestita Diels

    Directory of Open Access Journals (Sweden)

    Lanju Mei

    2017-09-01

    Conclusion: This work provides the first detailed transcriptome analysis of female and male flower of I. polycarpa and lays foundations for future studies on the molecular mechanisms underlying flower bud development of I. polycarpa.

  7. Ustawa budżetowa na 2016 rok – problematyka prawna

    OpenAIRE

    Borodo, Andrzej

    2016-01-01

    Ustawa budżetowa na 2016 r. ustala roczne dochody, wydatki, przychody, rozchody, koszty, wynagrodzenia, stawki należności, odpisy na fundusze. Ustalenia te dotyczą gospodarki finansowej rządu i innych części gospodarki finansowej państwa. Ustawa budżetowa zawiera postanowienia wynikające z wielu ustaw. Jest rozwinięciem lub wykonaniem uregulowań ujętych w całym ustawodawstwie. Ustawa budżetowa na 2016 r. zawiera trzy wymagane przez ustawę o finansach publicznych części: budżet, załączniki, in...

  8. Retained Cotton Bud-induced Severe Otitis Externa That Mimics Malignant Otitis Externa

    Directory of Open Access Journals (Sweden)

    Hyung Chae Yang

    2015-03-01

    Full Text Available The potential complications of cotton bud use are not well known in the elderly population. Here, we presented a case of cotton bud misuse. An elderly man who had long-standing diabetes mellitus, chronic otorrhea, and right-sided otalgia was poorly responsive to empirical treatment at a primary hospital. He was misdiagnosed with malignant otitis externa and referred to a tertiary hospital. On treatment Day 7 at the tertiary hospital, when the swollen ear canal was somewhat resolved, a retained cotton swab was found in his ear canal. His symptoms rapidly resolved after removing the retained foreign body. A cotton bud can be retained as a foreign body in the ear canal. The guidance of health professionals is needed to clean the ear canal using cotton buds, especially for elderly patients with other chronic diseases.

  9. Breast bud detection: a validation study in the Chilean growth obesity cohort study.

    Science.gov (United States)

    Pereira, Ana; Garmendia, María Luisa; González, Daniela; Kain, Juliana; Mericq, Verónica; Uauy, Ricardo; Corvalán, Camila

    2014-08-13

    Early puberty onset has been related to future chronic disease; however breast bud assessment in large scale population studies is difficult because it requires trained personnel. Thus our aim is to assess the validity of self and maternal breast bud detection, considering girl's body mass index (BMI) and maternal education. In 2010, 481 girls (mean age = 7.8) from the Growth and Obesity Chilean Cohort Study were evaluated by a nutritionist trained in breast bud detection. In addition, the girl(n = 481) and her mother(n = 341) classified the girl's breast development after viewing photographs of Tanner stages. Concordance between diagnostics was estimated (kappa, Spearman correlation) considering girls' BMI and mother's educational level. 14% of the girls presented breast buds and 43% had excess weight (BMI z-score > 1, World Health Organization 2007). Self-assessment showed low concordance with the evaluator (K puberty onset in epidemiological studies, particularly developing countries.

  10. Isolation and Characterization of Sesquiterpenoids from Cassia Buds and Their Antimicrobial Activities.

    Science.gov (United States)

    Guoruoluo, Yindengzhi; Zhou, Haofeng; Zhou, Junfei; Zhao, Haiqing; Aisa, Haji Akber; Yao, Guangmin

    2017-07-19

    Cassia buds, the immature fruits of Cinnamomum cassia (Lauraceae), are widely consumed as a food spice, dietary supplements, flavoring agents, and preservatives. In this study, cassia buds were phytochemically investigated for the first time, leading to the isolation of 2 new sesquiterpenoids (1 and 2) and 10 known sesquiterpenoids (3-12). Their structures were determined by spectrometric and spectroscopic analyses, including nuclear magnetic resonance, high-resolution electrospray ionization mass spectrometry, and circular dichroism. Cinnamosim A (1) represents the ninth example of the rare cyperane-type sesquiterpenoids. All of the compounds (1-12) isolated from cassia buds were evaluated for antimicrobial activities, with compounds 1-3, 5-8, 11, and 12 exhibiting strong antimicrobial activities against Candida albicans and compounds 6, 7, and 11 showing moderate antibacterial activities against Escherichia coli and Staphylococcus aureus. The present investigation indicated that sesquiterpenoids from cassia buds might be used as potential antimicrobial agents to preserve food.

  11. Identification of New Genes that Regulate Telomerase and Telomere Length in Budding Yeast

    National Research Council Canada - National Science Library

    Otero, Joel

    2003-01-01

    In budding yeast, Cdc13 has both an essential function in chromosome end protection as well as a non-essential role in telomere replication, by mediating recruitment of telomerase to the chromosome end...

  12. Karyotypic Determinants of Chromosome Instability in Aneuploid Budding Yeast

    Science.gov (United States)

    Bradford, William D.; Li, Rong

    2012-01-01

    Recent studies in cancer cells and budding yeast demonstrated that aneuploidy, the state of having abnormal chromosome numbers, correlates with elevated chromosome instability (CIN), i.e. the propensity of gaining and losing chromosomes at a high frequency. Here we have investigated ploidy- and chromosome-specific determinants underlying aneuploidy-induced CIN by observing karyotype dynamics in fully isogenic aneuploid yeast strains with ploidies between 1N and 2N obtained through a random meiotic process. The aneuploid strains exhibited various levels of whole-chromosome instability (i.e. chromosome gains and losses). CIN correlates with cellular ploidy in an unexpected way: cells with a chromosomal content close to the haploid state are significantly more stable than cells displaying an apparent ploidy between 1.5 and 2N. We propose that the capacity for accurate chromosome segregation by the mitotic system does not scale continuously with an increasing number of chromosomes, but may occur via discrete steps each time a full set of chromosomes is added to the genome. On top of such general ploidy-related effect, CIN is also associated with the presence of specific aneuploid chromosomes as well as dosage imbalance between specific chromosome pairs. Our findings potentially help reconcile the divide between gene-centric versus genome-centric theories in cancer evolution. PMID:22615582

  13. A taste for ATP: neurotransmission in taste buds

    Science.gov (United States)

    Kinnamon, Sue C.; Finger, Thomas E.

    2013-01-01

    Not only is ATP a ubiquitous source of energy but it is also used widely as an intercellular signal. For example, keratinocytes release ATP in response to numerous external stimuli including pressure, heat, and chemical insult. The released ATP activates purinergic receptors on nerve fibers to generate nociceptive signals. The importance of an ATP signal in epithelial-to-neuronal signaling is nowhere more evident than in the taste system. The receptor cells of taste buds release ATP in response to appropriate stimulation by tastants and the released ATP then activates P2X2 and P2X3 receptors on the taste nerves. Genetic ablation of the relevant P2X receptors leaves an animal without the ability to taste any primary taste quality. Of interest is that release of ATP by taste receptor cells occurs in a non-vesicular fashion, apparently via gated membrane channels. Further, in keeping with the crucial role of ATP as a neurotransmitter in this system, a subset of taste cells expresses a specific ectoATPase, NTPDase2, necessary to clear extracellular ATP which otherwise will desensitize the P2X receptors on the taste nerves. The unique utilization of ATP as a key neurotransmitter in the taste system may reflect the epithelial rather than neuronal origins of the receptor cells. PMID:24385952

  14. A comprehensive model to predict mitotic division in budding yeasts.

    Science.gov (United States)

    Sutradhar, Sabyasachi; Yadav, Vikas; Sridhar, Shreyas; Sreekumar, Lakshmi; Bhattacharyya, Dibyendu; Ghosh, Santanu Kumar; Paul, Raja; Sanyal, Kaustuv

    2015-11-05

    High-fidelity chromosome segregation during cell division depends on a series of concerted interdependent interactions. Using a systems biology approach, we built a robust minimal computational model to comprehend mitotic events in dividing budding yeasts of two major phyla: Ascomycota and Basidiomycota. This model accurately reproduces experimental observations related to spindle alignment, nuclear migration, and microtubule (MT) dynamics during cell division in these yeasts. The model converges to the conclusion that biased nucleation of cytoplasmic microtubules (cMTs) is essential for directional nuclear migration. Two distinct pathways, based on the population of cMTs and cortical dyneins, differentiate nuclear migration and spindle orientation in these two phyla. In addition, the model accurately predicts the contribution of specific classes of MTs in chromosome segregation. Thus we present a model that offers a wider applicability to simulate the effects of perturbation of an event on the concerted process of the mitotic cell division. © 2015 Sutradhar, Yadav, Sridhar, et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

  15. Timing robustness in the budding and fission yeast cell cycles.

    KAUST Repository

    Mangla, Karan

    2010-02-01

    Robustness of biological models has emerged as an important principle in systems biology. Many past analyses of Boolean models update all pending changes in signals simultaneously (i.e., synchronously), making it impossible to consider robustness to variations in timing that result from noise and different environmental conditions. We checked previously published mathematical models of the cell cycles of budding and fission yeast for robustness to timing variations by constructing Boolean models and analyzing them using model-checking software for the property of speed independence. Surprisingly, the models are nearly, but not totally, speed-independent. In some cases, examination of timing problems discovered in the analysis exposes apparent inaccuracies in the model. Biologically justified revisions to the model eliminate the timing problems. Furthermore, in silico random mutations in the regulatory interactions of a speed-independent Boolean model are shown to be unlikely to preserve speed independence, even in models that are otherwise functional, providing evidence for selection pressure to maintain timing robustness. Multiple cell cycle models exhibit strong robustness to timing variation, apparently due to evolutionary pressure. Thus, timing robustness can be a basis for generating testable hypotheses and can focus attention on aspects of a model that may need refinement.

  16. Initiation of Begonia erythrophylla L. vitroculture from axillary buds

    Directory of Open Access Journals (Sweden)

    Julieta - Emilia ROMOCEA

    2010-11-01

    Full Text Available In our aim was to establish a short-term in vitro culture of Begonia erythrophylla L., different culture media compositions were tested: complex variants added with growth regulators and simplified modified media with Heller microelements and MS macroelements. Adventitious shoots elongating over 13 mm in length were efficiently obtained from axillary bud segments of the strain of begonia Begonia erythrophylla L. on MS basic mineral medium culture containing different concentration of growth regulators, in order to identify optimal culture conditions, which would facilitate the achievement of a vitroculture, allowing the in vitro culture of this studied species. On complex regeneration variant V1 – mineral basic medium culture MB - MS supplemented with 1 mg/l BA, the rooting process was absent, but according to this carried out research, plantlets were obtained by rooting the elongated shoots on Murashige-Skoog (1962 basic mineral media containing 1 mg/l IBA, respectively a basic mineral medium culture without growth regulators, producing a much better organogenesis, where the phenomenon was greater in rooting process.

  17. The Genetic Requirements for Pentose Fermentation in Budding Yeast

    Directory of Open Access Journals (Sweden)

    Karin Mittelman

    2017-06-01

    Full Text Available Cells grow on a wide range of carbon sources by regulating substrate flow through the metabolic network. Incoming sugar, for example, can be fermented or respired, depending on the carbon identity, cell type, or growth conditions. Despite this genetically-encoded flexibility of carbon metabolism, attempts to exogenously manipulate central carbon flux by rational design have proven difficult, suggesting a robust network structure. To examine this robustness, we characterized the ethanol yield of 411 regulatory and metabolic mutants in budding yeast. The mutants showed little variation in ethanol productivity when grown on glucose or galactose, yet diversity was revealed during growth on xylulose, a rare pentose not widely available in nature. While producing ethanol at high yield, cells grown on xylulose produced ethanol at high yields, yet induced expression of respiratory genes, and were dependent on them. Analysis of mutants that affected ethanol productivity suggested that xylulose fermentation results from metabolic overflow, whereby the flux through glycolysis is higher than the maximal flux that can enter respiration. We suggest that this overflow results from a suboptimal regulatory adjustment of the cells to this unfamiliar carbon source.

  18. Characterization of Natural, Decellularized and Reseeded Porcine Tooth Bud Matrices

    Science.gov (United States)

    Traphagen, Samantha B.; Fourligas, Nikos; Xylas, Joanna; Sengupta, Sejuti; Kaplan, David; Georgakoudi, Irene; Yelick, Pamela C.

    2012-01-01

    Dental tissue engineering efforts have yet to identify scaffolds that instruct the formation of bioengineered teeth of predetermined size and shape. Here we investigated whether extracellular matrix (ECM) molecules present in natural tooth scaffolds can provide insight on how to achieve this goal. We describe methods to effectively decellularize and demineralize porcine molar tooth buds, while preserving natural ECM protein gradients. Natural tooth ECM composition was assessed using histological and immunohistochemical (IHC) analyses of fibrillar and basement membrane proteins. Our results showed that Collagen I, Fibronectin, Collagen IV, and Laminin gradients were detected in natural tooth tissues, and retained in decellularized samples. Second harmonic generation (SHG) image analysis and 3D reconstructions were used to show that natural tooth tissue exhibited higher collagen fiber density, and less oriented and less organized collagen fibers, as compared to decellularized tooth tissue. We also found that reseeded decellularized tooth scaffolds exhibited distinctive collagen content and organization as compared to decelluarized scaffolds. Our results show that SHG allows for quantitative assessment of ECM features that are not easily characterized using traditional histological analyses. In summary, our results demonstrate the potential for natural decellularized molar tooth ECM to instruct dental cell matrix synthesis, and lay the foundation for future use of biomimetic scaffolds for dental tissue engineering applications. PMID:22551485

  19. Partial repair of salinity-induced damage to sprouting sugarcane buds by proline and glycinebetaine pretreatment.

    Science.gov (United States)

    Rasheed, Rizwan; Wahid, Abdul; Hussain, Iqbal; Mahmood, Saqib; Parveen, Abida

    2016-05-01

    Sugarcane shows reduced crop stand under relatively suboptimal conditions; the main reason for this is its sensitivity to ionic stress in the soil solution. This research was performed to explore some physiological and developmental changes in the immature sugarcane buds submitted to salt stress and possible role of glycinebetaine (GB) and proline (Pro) in mitigating the ion toxicity in a time course manner. Salinity stress reduced fresh and dry weight, induced the generation of hydrogen peroxide, increased tissue levels of Na(+) sand Cl(-), reduced K(+) and Ca(2+), and K(+):Na(+) and Ca(2+):Na(+) ratios, while increasing the osmolyte synthesis in expanding sugarcane buds. Salinity stress reduced and delayed the formation of new bud leaves and their expansion, which was mainly because of reduction in the number and area of mesophyll cells and poor development of vascular bundles. The pretreatment of bud chips with 20 mM each of GB and Pro decreased tissue levels of Na(+) and Cl(-), reduced the generation of H2O2, improved K(+) and Ca(2+), K(+):Na(+) and Ca(2+):Na(+) ratios, and further increased the levels of GB, free proline (FP), and soluble sugars in the buds. The pretreatment increased mesophyll cell number and expansion of bud leaves and formation of elaborated vascular tissues, which apparently enabled the sprouting buds to adapt to salinity stress. Of the two osmolytes, GB was a relatively better inducer of salinity tolerance than Pro. In short, salinity-induced oxidative stress was the main cause for altered tissue development, the production of which was offset by pretreatment of bud tissues with Pro and GB.

  20. Glutamate may be an efferent transmitter that elicits inhibition in mouse taste buds.

    Directory of Open Access Journals (Sweden)

    Yijen A Huang

    Full Text Available Recent studies suggest that l-glutamate may be an efferent transmitter released from axons innervating taste buds. In this report, we determined the types of ionotropic synaptic glutamate receptors present on taste cells and that underlie this postulated efferent transmission. We also studied what effect glutamate exerts on taste bud function. We isolated mouse taste buds and taste cells, conducted functional imaging using Fura 2, and used cellular biosensors to monitor taste-evoked transmitter release. The findings show that a large fraction of Presynaptic (Type III taste bud cells (∼50% respond to 100 µM glutamate, NMDA, or kainic acid (KA with an increase in intracellular Ca(2+. In contrast, Receptor (Type II taste cells rarely (4% responded to 100 µM glutamate. At this concentration and with these compounds, these agonists activate glutamatergic synaptic receptors, not glutamate taste (umami receptors. Moreover, applying glutamate, NMDA, or KA caused taste buds to secrete 5-HT, a Presynaptic taste cell transmitter, but not ATP, a Receptor cell transmitter. Indeed, glutamate-evoked 5-HT release inhibited taste-evoked ATP secretion. The findings are consistent with a role for glutamate in taste buds as an inhibitory efferent transmitter that acts via ionotropic synaptic glutamate receptors.

  1. In vivo and ex vivo methods of growing a liver bud through tissue connection.

    Science.gov (United States)

    Yanagi, Yusuke; Nakayama, Koichi; Taguchi, Tomoaki; Enosawa, Shin; Tamura, Tadashi; Yoshimaru, Koichiro; Matsuura, Toshiharu; Hayashida, Makoto; Kohashi, Kenichi; Oda, Yoshinao; Yamaza, Takayoshi; Kobayashi, Eiji

    2017-10-26

    Cell-based therapy has been proposed as an alternative to orthotopic liver transplantation. The novel transplantation of an in vitro-generated liver bud might have therapeutic potential. In vivo and ex vivo methods for growing a liver bud are essential for paving the way for the clinical translation of liver bud transplantation. We herein report a novel transplantation method for liver buds that are grown in vivo involving orthotopic transplantation on the transected parenchyma of the liver, which showed long engraftment and marked growth in comparison to heterotopic transplantation. Furthermore, this study demonstrates a method for rapidly fabricating scalable liver-like tissue by fusing hundreds of liver bud-like spheroids using a 3D bioprinter. Its system to fix the shape of the 3D tissue with the needle-array system enabled the fabrication of elaborate geometry and the immediate execution of culture circulation after 3D printing-thereby avoiding an ischemic environment ex vivo. The ex vivo-fabricated human liver-like tissue exhibited self-tissue organization ex vivo and engraftment on the liver of nude rats. These achievements conclusively show both in vivo and ex vivo methods for growing in vitro-generated liver buds. These methods provide a new approach for in vitro-generated liver organoids transplantation.

  2. Circulating cytokeratin-positive cells and tumor budding in colorectal cancer.

    Science.gov (United States)

    Märkl, Bruno; Wilhelms, Narjes; Anthuber, Matthias; Schenkirsch, Gerhard; Schlimok, Günter; Oruzio, Daniel

    2016-12-10

    To investigate whether circulating cytokeratin-positive (CK(+)) cells in the mesenteric blood of resected colorectal specimens are prognostic and correlate with tumor budding. Fifty-six colorectal specimens were collected between 9/2007 and 7/2008. Blood from the mesenteric vein was drawn immediately after receiving the fresh and unfixed specimens in the pathology department. After separation of the mononuclear cells by Ficoll-Hypaque density-gradient centrifugation, cytological smears were immunocytochemically stained for CK18. Tumor budding was evaluated on slides stained for pan-cytokeratin. The identification of ≥ 30 buds/1.3 mm(2) was defined as high grade budding. CK(+) cells and clusters were identified in 29 (48%) and 14 (25%) of the samples, respectively. Two cells were identified in one of three non-malignant cases. Clusters were found exclusively in malignant cases. The occurrence of CK(+) cells or clusters was not associated with any of the evaluated clinicopathological factors, including surgical technique and tumor budding. Moreover, the occurrence of CK(+) cells or clusters had no influence on the cancer-specific survival [75 mo (CI: 61; 88) vs 83 mo (CI: 72; 95) and 80 mo (CI: 63; 98) vs 79 mo (CI: 69; 89), respectively]. CK(+) cells and showed neither prognostic significance nor an association with tumor budding. It is very likely that CK18-staining is not specific enough to identify the relevant cells.

  3. Tumor Budding in Intestinal Type Gastric Adenocarcinoma is Associated with Nodal Metastasis and Recurrence.

    Science.gov (United States)

    Olsen, Stephen; Linda, Jin; Fields, Ryan C; Yan, Yan; Nalbantoglu, ILKe

    2017-04-17

    Gastric adenocarcinoma (GAC) is a common cause of cancer-related death worldwide. GAC can be classified as intestinal or diffuse. Intestinal type cancers are common and reported to have a better prognosis compared to diffuse cancers. Studies have shown the presence and amount of tumor budding in intestinal carcinomas of the colon and esophagus to predict nodal metastasis and recurrence. Our aim is to determine if tumor budding in intestinal type GAC correlates with prognostic features. One hundred four patients treated with primary surgical excision between 1999-2013 were identified. Histologic type (intestinal, diffuse, or mixed), tumor grade, T-stage, and lymph node status were evaluated. Tumor bud scores were assigned to all intestinal type cancers using methods previously described for colorectal adenocarcinoma. Scores of Tumor characteristics were as follows: 52 intestinal (50%), 36 diffuse (35%) and 16 mixed (15%). Of the 52 cases with intestinal histology, 4 were well (8%), 28 were moderately (54%), and 20 were poorly differentiated (38%). Thirty-three (63%) of the intestinal tumors had high tumor bud scores. Cases with high scores were associated with higher T-stage, N-stage, and grade (Ptumor bud scores in intestinal type GAC have higher T-stage, N-stage, grade, and likelihood of recurrence. Assessment of tumor budding may guide clinical management in a subset of patients. Copyright © 2017. Published by Elsevier Inc.

  4. High-Grade Tumor Budding Stratifies Early-Stage Cervical Cancer with Recurrence Risk.

    Science.gov (United States)

    Huang, Bangxing; Cai, Jing; Xu, Xia; Guo, Shuang; Wang, Zehua

    2016-01-01

    This study investigated prognostic significance of tumor budding in early-stage cervical cancer (ESCC) following radical surgery and its contribution to improve the stratification of patients with recurrence risk. The archival medical records and H&E-stained slides of 643 patients with IA2-IIA stage cervical cancer who underwent radical surgery were retrospectively reviewed. Clinicopathological parameters were noted, and tumor buds were counted using immunohistochemistry for each case. The prognostic significance of tumor budding was analyzed. Prediction models that comprised tumor budding were established, and the performance was compared between the novel models and classic criteria via log-rank test and receiver operating characteristic analysis. Tumors with high-grade tumor budding (HTB) exhibited a substantially increased risk of recurrence (hazard ratio = 4.287, P tumor size ≥ 4 cm, deep stromal invasion of outer 1/3, and lymphovascular space invasion to stratify patients with an intermediate risk was most predictive of recurrence compared with the classic criteria. Tumor budding is an independent, unfavorable, prognostic factor for ESCC patients following radical surgery and holds promise for improved recurrence risk stratification.

  5. Tumor Budding Is Independently Predictive for Lymph Node Involvement in Early Gastric Cancer.

    Science.gov (United States)

    Gulluoglu, Mine; Yegen, Gulcin; Ozluk, Yasemin; Keskin, Metin; Dogan, Serap; Gundogdu, Gökçen; Onder, Semen; Balik, Emre

    2015-08-01

    The most important prognostic factor for early gastric cancer (EGC) is the lymph node status. It is important to predict early lesions without lymph node metastasis (LNM) before proceeding to radical surgery in locally excised lesions. Tumor budding is a feature known to be related to aggressive tumor behavior in several solid tumors. We aimed to assess the predictive value of tumor budding for LNM in pT1a and pT1b gastric cancer. We retrospectively investigated radical gastrectomy specimens for of 126 EGC patients and assess the possible relation between the clinicopathologic features, including age, gender, tumor location, tumor size, macroscopic tumor type, histologic differentiation, depth and width of submucosal invasion, lymphovascular invasion, and tumor budding with lymph node involvement. Among the 126 EGCs, 38 were stages as pT1a and 88 as pT1b. LNM rate in pT1a tumors was 13% whereas it was 33% in pT1b tumors. Tumor budding was the only factor significantly and independently related to LNM in pT1a patients. Female gender and tumor budding were found to be independent risk factors in pT1b group. Other clinicopathologic features were not related to LNM. Based on these results, we suggest that budding is a promising parameter to assess for prediction of LNM in EGC removed by endoscopic surgery, and to decide on the appropriate surgical approach. © The Author(s) 2015.

  6. Identification of genes associated with bud dormancy release in Prunus persica by suppression subtractive hybridization.

    Science.gov (United States)

    Leida, Carmen; Terol, Javier; Martí, Gracia; Agustí, Manuel; Llácer, Gerardo; Badenes, María Luisa; Ríos, Gabino

    2010-05-01

    To better understand the molecular and physiological mechanisms underlying maintenance and release of seasonal bud dormancy in perennial trees, we identified differentially expressed genes during dormancy progression in reproductive buds from peach (Prunus persica [L.] Batsch) by suppression subtractive hybridization (SSH) and microarray hybridization. Four SSH libraries were constructed, which were respectively enriched in cDNA highly expressed in dormant buds (named DR), in dormancy-released buds (RD) and in the cultivars with different chilling requirement, 'Zincal 5' (ZS) and 'Springlady' (SZ), sampled after dormancy release. About 2500 clones picked from the four libraries were loaded on a glass microarray. Hybridization of microarrays with the final products of SSH procedure was performed in order to validate the selected clones that were effectively enriched in their respective sample. Nearly 400 positive clones were sequenced, which corresponded to 101 different unigenes with diverse functional annotation. We obtained DAM4, 5 and 6 genes coding for MADS-box transcription factors previously related to growth cessation and terminal bud formation in the evergrowing mutant of peach. Several other cDNAs are similar to dormancy factors described in other species, and others have been related to bud dormancy for the first time in this study. Quantitative reverse transcription polymerase chain reaction analysis confirmed differential expression of cDNAs coding for a Zn-finger transcription factor, a GRAS-like regulator, a DNA-binding protein and proteins similar to forisome subunits involved in the reversible occlusion of sieve elements in Fabaceae, among others.

  7. [Research of the amounts of flavonoids accumulated in the buds of single-styled hawthorn].

    Science.gov (United States)

    Jakstas, Valdas; Janulis, Valdimaras; Labokas, Juozas

    2004-01-01

    To determine the amounts of flavonoids accumulated in the buds of single-styled hawthorn (Crataegus monogyna Jacq.); and to establish the possibility of usage of hawthorn buds in pharmacy practice. Different examples of hawthorn buds collected in 2001, 2002 and 2003 from some Lithuanian regions: Akmene (V1, V2, V3), Kedainiai (S1, S2, S3), Klaipeda (M1), Lazdijai (D1, D2, D3) and Vilnius (VL1, VL2) from branches of lower storey. Examples were extracted with ethanol and flavonoids were measured by spectroscopic method and by high performance liquid chromatography. From 0.72 to 1.89% of flavonoids are found in the hawthorn buds. The dominating flavonoid is vitexin-2-O-rhamnosid (6.72-10.91 milligrams in one gram of dried crude drug). Other flavonoids are: vitexin: 0.88-6.53 milligrams/gram, hyperosid: 0.85-2.70 milligrams/gram, rutin: 0.72-2.10 milligrams/gram and quercitrin: 0.82-1.01 milligrams/gram. The marks of phytochemical compounds of Lithuanian single-styled hawthorn buds corroborated the theory of possibility to use hawthorn buds as pharmaceutical crude drug and form sufficient ground for planning phytochemical and pharmacological researches of new hawthorn crude drug.

  8. Cell morphology, budding propensity and cell death of Saccharomyces cerevisiae at high hydrostatic pressure

    Science.gov (United States)

    Nguyen, Khanh; Lewis, Jeffrey; Kumar, Pradeep

    A large biomass on earth thrives in extremes of physical and chemical conditions including high pressure and temperature. Budding yeast, S. cerevisiae, is a eukaryotic model organism due to its amenability to molecular biology tools. To understand the effects of hydrostatic pressure on a eukaryotic cell, we have performed quantitative experiments of the growth, the propensity of budding, and cell death of S. cerevisiae in a wide range of pressures. An automated image analysis method for the quantification of the budding index was developed and applied along with a continuum model of budding to investigate the effects of pressure on cell division and cell morphology. We find that the growth, the budding propensity, the average cell size, and the ellipticity of the cells decrease with increasing pressure. Furthermore, large hydrostatic pressure led to the small but finite probability of cell death. Our experiments suggest that the decrease of budding propensity arises from cellular arrest at the cell cycle checkpoints during different stages of cell division.

  9. Effect of static magnetic fields on the budding of yeast cells.

    Science.gov (United States)

    Egami, Shigeki; Naruse, Yujiro; Watarai, Hitoshi

    2010-12-01

    The effect of static magnetic fields on the budding of single yeast cells was investigated using a magnetic circuit that was capable of generating a strong magnetic field (2.93 T) and gradient (6100 T²  m⁻¹). Saccharomyces cerevisiae yeast cells were grown in an aqueous YPD agar in a silica capillary under either a homogeneous or inhomogeneous static magnetic field. Although the size of budding yeast cells was only slightly affected by the magnetic fields after 4 h, the budding angle was clearly affected by the direction of the homogeneous and inhomogeneous magnetic fields. In the homogeneous magnetic field, the budding direction of daughter yeast cells was mainly oriented in the direction of magnetic field B. However, when subjected to the inhomogeneous magnetic field, the daughter yeast cells tended to bud along the axis of capillary flow in regions where the magnetic gradient, estimated by B(dB/dx), were high. Based on the present experimental results, the possible mechanism for the magnetic effect on the budding direction of daughter yeast cells is theoretically discussed. Copyright © 2010 Wiley-Liss, Inc.

  10. Influence of the bud neck on nuclear envelope fission in Saccharomyces cerevisiae.

    Science.gov (United States)

    Melloy, Patricia G; Rose, Mark D

    2017-09-15

    Studies have shown that nuclear envelope fission (karyokinesis) in budding yeast depends on cytokinesis, but not distinguished whether this was a direct requirement, indirect, because of cell cycle arrest, or due to bud neck-localized proteins impacting both processes. To determine the requirements for karyokinesis, we examined mutants conditionally defective for bud emergence and/or nuclear migration. The common mutant phenotype was completion of the nuclear division cycle within the mother cell, but karyokinesis did not occur. In the cdc24 swe1 mutant, at the non-permissive temperature, multiple nuclei accumulated within the unbudded cell, with connected nuclear envelopes. Upon return to the permissive temperature, the cdc24 swe1 mutant initiated bud emergence, but only the nucleus spanning the neck underwent fission suggesting that the bud neck region is important for fission initiation. The neck may be critical for either mechanical reasons, as the contractile ring might facilitate fission, or for regulatory reasons, as the site of a protein network regulating nuclear envelope fission, mitotic exit, and cytokinesis. We also found that 77-85% of pairs of septin mutant nuclei completed nuclear envelope fission. In addition, 27% of myo1Δ mutant nuclei completed karyokinesis. These data suggested that fission is not dependent on mechanical contraction at the bud neck, but was instead controlled by regulatory proteins there. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. Triterpenoid saponins from the buds of Lonicera similis.

    Science.gov (United States)

    Zhang, Xiao; Zou, Li-Hua; He, Yu-Lin; Peng, Cheng; Guo, Li; Xiong, Liang

    2017-11-30

    Four new lupane triterpenoid saponins, along with one known lupane and eight hederagenin saponins, were isolated from the EtOH extract of the buds of Lonicera similis Hemsl. The structures of the new compounds were established as 3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl 23-hydroxybetulinic acid 28-O-β-D-glucopyranosyl ester (lonisimilioside A, 1), 3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl 23-hydroxybetulinic acid 28-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester (lonisimilioside B, 2), 3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl betulinic acid 28-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester (lonisimilioside C, 3) and 3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl betulinic acid 28-O-β-D-glucopyranosyl ester (lonisimilioside D, 4), respectively. The cytotoxic activities of the isolates against human cancer cell lines HepG2, MCF-7 and A-549 were evaluated. Only the monodesmosidic saponin with a free carboxyl group at C-28 (12) exhibited significant cytotoxicities against HepG2, MCF-7 and A-549 cell lines with the IC 50 values of 8.98 ± 0.19, 12.48 ± 0.45 and 11.62 ± 0.54 μM, respectively. Furthermore, Hoechst fluorescence 33342 staining was used to demonstrate that 12 could induce HepG2 and A-549 cells apoptosis significantly.

  12. Origin of irreversibility of cell cycle start in budding yeast.

    Directory of Open Access Journals (Sweden)

    Gilles Charvin

    2010-01-01

    Full Text Available Budding yeast cells irreversibly commit to a new division cycle at a regulatory transition called Start. This essential decision-making step involves the activation of the SBF/MBF transcription factors. SBF/MBF promote expression of the G1 cyclins encoded by CLN1 and CLN2. Cln1,2 can activate their own expression by inactivating the Whi5 repressor of SBF/MBF. The resulting transcriptional positive feedback provides an appealing, but as yet unproven, candidate for generating irreversibility of Start. Here, we investigate the logic of the Start regulatory module by quantitative single-cell time-lapse microscopy, using strains in which expression of key regulators is efficiently controlled by changes of inducers in a microfluidic chamber. We show that Start activation is ultrasensitive to G1 cyclin. In the absence of CLN1,2-dependent positive feedback, we observe that Start transit is reversible, due to reactivation of the Whi5 transcriptional repressor. Introduction of the positive feedback loop makes Whi5 inactivation and Start activation irreversible, which therefore guarantees unidirectional entry into S phase. A simple mathematical model to describe G1 cyclin turn on at Start, entirely constrained by empirically measured parameters, shows that the experimentally measured ultrasensitivity and transcriptional positive feedback are necessary and sufficient dynamical characteristics to make the Start transition a bistable and irreversible switch. Our study thus demonstrates that Start irreversibility is a property that arises from the architecture of the system (Whi5/SBF/Cln2 loop, rather than the consequence of the regulation of a single component (e.g., irreversible protein degradation.

  13. Systems Level Modeling of the Cell Cycle Using Budding Yeast

    Directory of Open Access Journals (Sweden)

    D.R. Kim

    2007-01-01

    Full Text Available Proteins involved in the regulation of the cell cycle are highly conserved across all eukaryotes, and so a relatively simple eukaryote such as yeast can provide insight into a variety of cell cycle perturbations including those that occur in human cancer. To date, the budding yeast Saccharomyces cerevisiae has provided the largest amount of experimental and modeling data on the progression of the cell cycle, making it a logical choice for in-depth studies of this process. Moreover, the advent of methods for collection of high-throughput genome, transcriptome, and proteome data has provided a means to collect and precisely quantify simultaneous cell cycle gene transcript and protein levels, permitting modeling of the cell cycle on the systems level. With the appropriate mathematical framework and suffi cient and accurate data on cell cycle components, it should be possible to create a model of the cell cycle that not only effectively describes its operation, but can also predict responses to perturbations such as variation in protein levels and responses to external stimuli including targeted inhibition by drugs. In this review, we summarize existing data on the yeast cell cycle, proteomics technologies for quantifying cell cycle proteins, and the mathematical frameworks that can integrate this data into representative and effective models. Systems level modeling of the cell cycle will require the integration of high-quality data with the appropriate mathematical framework, which can currently be attained through the combination of dynamic modeling based on proteomics data and using yeast as a model organism.

  14. The cellular robustness by genetic redundancy in budding yeast.

    Directory of Open Access Journals (Sweden)

    Jingjing Li

    2010-11-01

    Full Text Available The frequent dispensability of duplicated genes in budding yeast is heralded as a hallmark of genetic robustness contributed by genetic redundancy. However, theoretical predictions suggest such backup by redundancy is evolutionarily unstable, and the extent of genetic robustness contributed from redundancy remains controversial. It is anticipated that, to achieve mutual buffering, the duplicated paralogs must at least share some functional overlap. However, counter-intuitively, several recent studies reported little functional redundancy between these buffering duplicates. The large yeast genetic interactions released recently allowed us to address these issues on a genome-wide scale. We herein characterized the synthetic genetic interactions for ∼500 pairs of yeast duplicated genes originated from either whole-genome duplication (WGD or small-scale duplication (SSD events. We established that functional redundancy between duplicates is a pre-requisite and thus is highly predictive of their backup capacity. This observation was particularly pronounced with the use of a newly introduced metric in scoring functional overlap between paralogs on the basis of gene ontology annotations. Even though mutual buffering was observed to be prevalent among duplicated genes, we showed that the observed backup capacity is largely an evolutionarily transient state. The loss of backup capacity generally follows a neutral mode, with the buffering strength decreasing in proportion to divergence time, and the vast majority of the paralogs have already lost their backup capacity. These observations validated previous theoretic predictions about instability of genetic redundancy. However, departing from the general neutral mode, intriguingly, our analysis revealed the presence of natural selection in stabilizing functional overlap between SSD pairs. These selected pairs, both WGD and SSD, tend to have decelerated functional evolution, have higher propensities of co

  15. Differentiated dynamics of bud dormancy and growth in temperate fruit trees relating to bud phenology adaptation, the case of apple and almond trees

    Science.gov (United States)

    El Yaacoubi, Adnane; Malagi, Gustavo; Oukabli, Ahmed; Citadin, Idemir; Hafidi, Majida; Bonhomme, Marc; Legave, Jean-Michel

    2016-11-01

    Few studies have focused on the characterization of bud dormancy and growth dynamics for temperate fruit species in temperate and mild cropping areas, although this is an appropriate framework to anticipate phenology adaptation facing future warming contexts which would potentially combine chill declines and heat increases. To examine this issue, two experimental approaches and field observations were used for high- and low-chill apple cultivars in temperate climate of southern France and in mild climates of northern Morocco and southern Brazil. Low-chill almond cultivars offered an additional relevant plant material for comparison with apple in northern Morocco. Divergent patterns of dormancy and growth dynamics were clearly found in apple tree between southern France and southern Brazil. Divergences were less pronounced between France and Morocco. A global view outlined main differences in the dormancy chronology and intensity, the transition between endordormancy and ecodormancy and the duration of ecodormancy. A key role of bud rehydration in the transition period was shown. High-chill cultivars would be submitted in mild conditions to heterogeneous rehydration capacities linked to insufficient chill fulfillment and excessive forcing linked to high temperatures. This would favor bud competitions and consequently excessive flowering durations and weak flowering. Low chilling requirements in apple and almond would conversely confer biological capacities to tolerate superficial dormancy and abrupt transition from endordormancy to ecodormancy without important heterogeneous rehydration states within buds. It may also assume that low-chill cultivars can also tolerate high temperatures during ecodormancy as well as extended flowering durations.

  16. Local adaptations and climate change: converging sensitivity of bud break in black spruce provenances

    Science.gov (United States)

    Rossi, Sergio

    2015-07-01

    Species with transcontinental distribution or spread over wide geographical regions develop populations with growth traits genetically adapted to the local climate. The aim of this study was to investigate the ecotypic sensitivity of bud break, a strong adaptive trait, to a changing environment. Six phenological phases of bud break were monitored daily on black spruce [ Picea mariana (Mill.) BSP] seedlings submitted to different temperatures (12, 16 and 20 °C) and photoperiods (14, 18 and 22 h). Six provenances were tested in growth chambers, produced from seeds collected along the whole latitudinal range of the closed boreal forest in Quebec, Canada. Bud break lasted 13.3 days on average and occurred earlier in seedlings from colder sites. The annual temperature of the sites suitably tracked the clinal variation among ecotypes, providing a clear biological explanation for the environmental signal driving the adaptive divergence of populations to the local climate. Increasing temperature induced an earlier bud break according to a non-linear pattern with greater advancements observed between 12 and 16 °C. Photoperiod was significant, but sensitivity analysis indicated that its effect on bud break was marginal with respect to temperature. No interaction of provenance × treatment was observed, demonstrating an ecotypic convergence of the responses to both factors. Changes in the growing conditions could substantially modify the synchronization between bud phenology and climate, thus exposing the developing meristems of black spruce to frost damage. However, similar advancements of bud break could be expected in the different ecotypes subjected to warmer temperatures or longer day lengths.

  17. Bud development in corydalis (Corydalis bracteata) requires low temperature: a study of developmental and carbohydrate changes

    Science.gov (United States)

    Khodorova, Nadejda V.; Miroslavov, Evgeniy A.; Shavarda, Alexey L.; Laberche, Jean-Claude; Boitel-Conti, Michèle

    2010-01-01

    Background and Aims Spring geophytes require a period of low temperature for proper flower development but the mechanism that underlies the relationship between cold treatment and flowering remains unknown. The present study aims to compare the developmental anatomy and carbohydrate content of the tuberous geophyte Corydalis bracteata growing under natural winter conditions from 10 to −10 °C (field-grown) and under a mild temperature regime of 18 °C (indoor-grown plants). Methods Samples were studied under light and electron microscopy. A histochemical test (periodic acid – Schiff's) was employed to identify starch in sectioned material. Sugars were analysed by capillary gas chromatography. Apoplastic wash fluid was prepared. Key Results Under natural conditions, shoots were elongated, and buds gained in dry mass and developed normally. For indoor-grown plants, these parameters were lower in value and, from December, a progressive necrosis of flower buds was observed. The tuber consisted of the new developing one, which was connected to the bud, and the old tuber with its starch reserve. Due to the absence of plasmodesmata between new and old tuber cells, sugar transport cannot be through the symplast. Thus, a potential apoplastic route is proposed from old tuber phloem parenchyma cells to the adjacent new tuber cells. Sugar content in buds during the autumn months (September–November) was lower for indoor-grown plants than control plants, whereas the sugar content in tubers during the same period was similar for plants from both temperature treatments. However, the amount of apoplastic sugars in tubers of field-grown plants was almost 15-fold higher than in indoor-grown tubers. Conclusions The results suggest that low temperature activates the apoplastic route of sugar transport in C. bracteata tubers and a consequent carbohydrate delivery to the bud. In the absence of cold treatment, the carbohydrate reserve is locked in old tuber cells so the nutrient

  18. Functional interchangeability of late domains, late domain cofactors and ubiquitin in viral budding.

    Directory of Open Access Journals (Sweden)

    Maria Zhadina

    2010-10-01

    Full Text Available The membrane scission event that separates nascent enveloped virions from host cell membranes often requires the ESCRT pathway, which can be engaged through the action of peptide motifs, termed late (L- domains, in viral proteins. Viral PTAP and YPDL-like L-domains bind directly to the ESCRT-I and ALIX components of the ESCRT pathway, while PPxY motifs bind Nedd4-like, HECT-domain containing, ubiquitin ligases (e.g. WWP1. It has been unclear precisely how ubiquitin ligase recruitment ultimately leads to particle release. Here, using a lysine-free viral Gag protein derived from the prototypic foamy virus (PFV, where attachment of ubiquitin to Gag can be controlled, we show that several different HECT domains can replace the WWP1 HECT domain in chimeric ubiquitin ligases and drive budding. Moreover, artificial recruitment of isolated HECT domains to Gag is sufficient to stimulate budding. Conversely, the HECT domain becomes dispensable if the other domains of WWP1 are directly fused to an ESCRT-1 protein. In each case where budding is driven by a HECT domain, its catalytic activity is essential, but Gag ubiquitination is dispensable, suggesting that ubiquitin ligation to trans-acting proteins drives budding. Paradoxically, however, we also demonstrate that direct fusion of a ubiquitin moiety to the C-terminus of PFV Gag can also promote budding, suggesting that ubiquitination of Gag can substitute for ubiquitination of trans-acting proteins. Depletion of Tsg101 and ALIX inhibits budding that is dependent on ubiquitin that is fused to Gag, or ligated to trans-acting proteins through the action of a PPxY motif. These studies underscore the flexibility in the ways that the ESCRT pathway can be engaged, and suggest a model in which the identity of the protein to which ubiquitin is attached is not critical for subsequent recruitment of ubiquitin-binding components of the ESCRT pathway and viral budding to proceed.

  19. Modeling of bud break of Scots pine in northern Finland in 1908–2014

    Science.gov (United States)

    Salminen, Hannu; Jalkanen, Risto

    2015-01-01

    Bud break and height-growth of Scots pine (Pinus sylvestris L.) in the northern boreal zone in Lapland, Finland, was followed through the entire growing seasons in the periods 2001–2003 and 2008–2010 in sapling stands in two different locations in northern Finland set some 250 km apart along a latitudinal transect. Field measurements continued at the southern site also in 2011–2013. Air temperature was recorded hourly at the sites. A simple optimization algorithm (GA) was used to adjust parameters of the models predicting the timing of bud break of Scots pine in order to minimize the difference between observed and predicted dates. The models giving the best performance and century-long daily temperatures were used to reconstruct bud-break time series. The temperature observations were recorded for the period 1908–2014 in Sodankylä, which is located in-between the sapling stands in the north–south direction and for the period 1877–2014 in Karasjok, which is in Norway about 145 km north–west from the northernmost stand of this study. On average buds began to extend in the beginning of May in the southernmost stand and in mid-May in the northernmost stands, and the variation between years was in the range of 3 weeks. A simple day-length-triggered (fixed date) model predicted most accurately the date of bud break; root mean square error (RMSE) was 2 and 4 days in the northern and southern site, respectively. The reconstructed bud-break series indicated that based on temperature observations from Sodankylä, growth onset of Scots pine has clearly advanced since the 1960s, though it currently matches that of the early 1920s and early 1950s. The temperature record from Karasjok indicated a similar variation, though there was a weak linear trend advancing bud break by about 3–4 days over a 100-year period. PMID:25798141

  20. Modeling of bud break of Scots pine in northern Finland in 1908-2014.

    Science.gov (United States)

    Salminen, Hannu; Jalkanen, Risto

    2015-01-01

    Bud break and height-growth of Scots pine (Pinus sylvestris L.) in the northern boreal zone in Lapland, Finland, was followed through the entire growing seasons in the periods 2001-2003 and 2008-2010 in sapling stands in two different locations in northern Finland set some 250 km apart along a latitudinal transect. Field measurements continued at the southern site also in 2011-2013. Air temperature was recorded hourly at the sites. A simple optimization algorithm (GA) was used to adjust parameters of the models predicting the timing of bud break of Scots pine in order to minimize the difference between observed and predicted dates. The models giving the best performance and century-long daily temperatures were used to reconstruct bud-break time series. The temperature observations were recorded for the period 1908-2014 in Sodankylä, which is located in-between the sapling stands in the north-south direction and for the period 1877-2014 in Karasjok, which is in Norway about 145 km north-west from the northernmost stand of this study. On average buds began to extend in the beginning of May in the southernmost stand and in mid-May in the northernmost stands, and the variation between years was in the range of 3 weeks. A simple day-length-triggered (fixed date) model predicted most accurately the date of bud break; root mean square error (RMSE) was 2 and 4 days in the northern and southern site, respectively. The reconstructed bud-break series indicated that based on temperature observations from Sodankylä, growth onset of Scots pine has clearly advanced since the 1960s, though it currently matches that of the early 1920s and early 1950s. The temperature record from Karasjok indicated a similar variation, though there was a weak linear trend advancing bud break by about 3-4 days over a 100-year period.

  1. Local adaptations and climate change: converging sensitivity of bud break in black spruce provenances.

    Science.gov (United States)

    Rossi, Sergio

    2015-07-01

    Species with transcontinental distribution or spread over wide geographical regions develop populations with growth traits genetically adapted to the local climate. The aim of this study was to investigate the ecotypic sensitivity of bud break, a strong adaptive trait, to a changing environment. Six phenological phases of bud break were monitored daily on black spruce [Picea mariana (Mill.) BSP] seedlings submitted to different temperatures (12, 16 and 20 °C) and photoperiods (14, 18 and 22 h). Six provenances were tested in growth chambers, produced from seeds collected along the whole latitudinal range of the closed boreal forest in Quebec, Canada. Bud break lasted 13.3 days on average and occurred earlier in seedlings from colder sites. The annual temperature of the sites suitably tracked the clinal variation among ecotypes, providing a clear biological explanation for the environmental signal driving the adaptive divergence of populations to the local climate. Increasing temperature induced an earlier bud break according to a non-linear pattern with greater advancements observed between 12 and 16 °C. Photoperiod was significant, but sensitivity analysis indicated that its effect on bud break was marginal with respect to temperature. No interaction of provenance × treatment was observed, demonstrating an ecotypic convergence of the responses to both factors. Changes in the growing conditions could substantially modify the synchronization between bud phenology and climate, thus exposing the developing meristems of black spruce to frost damage. However, similar advancements of bud break could be expected in the different ecotypes subjected to warmer temperatures or longer day lengths.

  2. Bud sprouting and floral induction and expression of FT in loquat [Eriobotrya japonica (Thunb.) Lindl.].

    Science.gov (United States)

    Reig, Carmina; Gil-Muñoz, Francisco; Vera-Sirera, Francisco; García-Lorca, Ana; Martínez-Fuentes, Amparo; Mesejo, Carlos; Pérez-Amador, Miguel A; Agustí, Manuel

    2017-07-14

    EjFT1 and EjFT2 genes were isolated and sequenced from leaves of loquat. EjFT1 is involved in bud sprouting and leaf development, and EjFT2 in floral bud induction. Loquat [Eriobotrya japonica (Thunb.) Lindl.] is an evergreen species belonging to the family Rosaceae, such as apple and pear, whose reproductive development, in contrast with these species, is a continuous process that is not interrupted by winter dormancy. Thus, the study of the mechanism of flowering in loquat has the potential to uncover the environmental and genetic networks that trigger flowering more accurately, contributing for a better understanding of the Rosaceae floral process. As a first step toward understanding the molecular mechanisms controlling flowering, extensive defoliation and defruiting assays, together with molecular studies of the key FLOWERING LOCUS T (FT) gene, were carried out. FT exhibited two peaks of expression in leaves, the first one in early to mid-May, the second one in mid-June. Two FT genes, EjFT1 and EjFT2, were isolated and sequenced and studied their expression. Expression of EjFT1 and EjFT2 peaks in mid-May, at bud sprouting. EjFT2 expression peaks again in mid-June, coinciding with the floral bud inductive period. Thus, when all leaves of the tree were continuously removed from early to late May vegetative apex differentiated into panicle, but when defoliation was performed from early to late June apex did not differentiate. On the other hand, fruit removal advanced EjFT1 expression in old leaves and the sooner the fruit detached, the sooner the bud sprouted. Accordingly, results strongly suggest that EjFT1 might be related to bud sprouting and leaf development, while EjFT2 might be involved in floral bud induction. An integrative model for FT functions in loquat is discussed.

  3. 5'-end sequences of budding yeast full-length cDNA clones and quality scores - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available List Contact us Budding yeast cDNA sequencing project 5'-end sequences of budding yeast full-length cDNA clones and quality score...s Data detail Data name 5'-end sequences of budding yeast full-length cDNA clones and quality score...or-capping method, the sequence quality score generated by the Phred software, and links to SGD, dbEST and U...es. FASTA format. Quality Phred's quality score About This Database Database Desc...g yeast full-length cDNA clones and quality scores - Budding yeast cDNA sequencing project | LSDB Archive ...

  4. BUD31 and Lipid Metabolism: A New Potential Therapeutic Entry Point for Myc-Driven Breast Cancer

    Science.gov (United States)

    2016-02-01

    AWARD NUMBER: W81XWH-14-1-0039 TITLE: BUD31 and Lipid Metabolism: A New Potential Therapeutic Entry Point for Myc-Driven Breast Cancer...TITLE AND SUBTITLE 5a. CONTRACT NUMBER BUD31 and Lipid Metabolism: A New Potential Therapeutic Entry Point for Myc-Driven Breast Cancer 5b. GRANT...manuscript in Nature detailing a role for BUD31 in splicing and shown more broadly that splicing is a viable therapeutic intervention point for myc

  5. Tumor budding at the invasive front of colorectal cancer may not be associated with the epithelial-mesenchymal transition.

    Science.gov (United States)

    Yamada, Noriyuki; Sugai, Tamotsu; Eizuka, Makoto; Tsuchida, Koudai; Sugimoto, Ryo; Mue, Yoshiharu; Suzuki, Masamichi; Osakabe, Mitsumasa; Uesugi, Noriyuki; Ishida, Kazuyuki; Otsuka, Kouki; Matsumoto, Takayuki

    2017-02-01

    Tumor budding is thought to reflect the epithelial-mesenchymal transition (EMT). However, the molecular mechanism linking tumor buds and the EMT remains unclear. Here, we examined the induction of tumor budding and EMT and their association with EMT-related proteins (ZEB1, TWIST, SNAIL, and SLUG) in colorectal cancer (CRC). Immunohistochemical expression of pan-cytokeratin was examined for identification of tumor budding in 101 CRCs. Grading of tumor budding was classified into low- and high-grade groups. Tissue microarray was conducted to identify tumor budding sites. The expression of E-cadherin, ZEB1, TWIST, SNAIL, and SLUG was examined in areas of tumor budding and the surrounding tumor stroma using a double-immunostaining method. Specifically, pan-cytokeratin and EMT-related proteins were assessed by double immunostaining. Low or no expression of E-cadherin was found in areas of tumor budding. Moreover, ZEB1, TWIST, SNAIL, and SLUG were not expressed in regions of tumor budding. However, the expression level of ZEB1 in the stromal cells surrounding tumor budding was significantly more frequent than that of TWIST, SNAI, and SLUG. In addition, the expression of EMT-related proteins in surrounding stromal cells was significantly greater in areas of high-grade tumor budding than in low-grade areas. Our present results suggest that EMT-related proteins play a minor role in forming tumor buds. In addition, our findings suggest the existence of subtypes of stromal cells in CRC with phenotypical and functional heterogeneity. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Spatio-temporal relief from hypoxia and production of reactive oxygen species during bud burst in grapevine (Vitis vinifera).

    Science.gov (United States)

    Meitha, Karlia; Konnerup, Dennis; Colmer, Timothy D; Considine, John A; Foyer, Christine H; Considine, Michael J

    2015-09-01

    Plants regulate cellular oxygen partial pressures (pO2), together with reduction/oxidation (redox) state in order to manage rapid developmental transitions such as bud burst after a period of quiescence. However, our understanding of pO2 regulation in complex meristematic organs such as buds is incomplete and, in particular, lacks spatial resolution. The gradients in pO2 from the outer scales to the primary meristem complex were measured in grapevine (Vitis vinifera) buds, together with respiratory CO2 production rates and the accumulation of superoxide and hydrogen peroxide, from ecodormancy through the first 72 h preceding bud burst, triggered by the transition from low to ambient temperatures. Steep internal pO2 gradients were measured in dormant buds with values as low as 2·5 kPa found in the core of the bud prior to bud burst. Respiratory CO2 production rates increased soon after the transition from low to ambient temperatures and the bud tissues gradually became oxygenated in a patterned process. Within 3 h of the transition to ambient temperatures, superoxide accumulation was observed in the cambial meristem, co-localizing with lignified cellulose associated with pro-vascular tissues. Thereafter, superoxide accumulated in other areas subtending the apical meristem complex, in the absence of significant hydrogen peroxide accumulation, except in the cambial meristem. By 72 h, the internal pO2 gradient showed a biphasic profile, where the minimum pO2 was external to the core of the bud complex. Spatial and temporal control of the tissue oxygen environment occurs within quiescent buds, and the transition from quiescence to bud burst is accompanied by a regulated relaxation of the hypoxic state and accumulation of reactive oxygen species within the developing cambium and vascular tissues of the heterotrophic grapevine buds. © The Author 2015. Published by Oxford University Press on behalf of the Annals of Botany Company.

  7. Cytokinins and Abscisic Acid Act Antagonistically in the Regulation of the Bud Outgrowth Pattern by Light Intensity

    Directory of Open Access Journals (Sweden)

    Adrien Corot

    2017-10-01

    Full Text Available Bud outgrowth is a key process in the elaboration of yield and visual quality in rose crops. Although light intensity is well known to affect bud outgrowth, little is known on the mechanisms involved in this regulation. The objective of this work was to test if the control of bud outgrowth pattern along the stem by photosynthetic photon flux density (PPFD is mediated by sugars, cytokinins and/or abscisic acid in intact rose plants. Rooted cuttings of Rosa hybrida ‘Radrazz’ were grown in growth chambers under high PPFD (530 μmol m-2 s-1 until the floral bud visible stage. Plants were then either placed under low PPFD (90 μmol m-2 s-1 or maintained under high PPFD. Bud outgrowth inhibition by low PPFD was associated with lower cytokinin and sugar contents and a higher abscisic acid content in the stem. Interestingly, cytokinin supply to the stem restored bud outgrowth under low PPFD. On the other hand, abscisic acid supply inhibited outgrowth under high PPFD and antagonized bud outgrowth stimulation by cytokinins under low PPFD. In contrast, application of sugars did not restore bud outgrowth under low PPFD. These results suggest that PPFD regulation of bud outgrowth in rose involves a signaling pathway in which cytokinins and abscisic acid play antagonistic roles. Sugars can act as nutritional and signaling compounds and may be involved too, but do not appear as the main regulator of the response to PPFD.

  8. Antioxidant effects of clove bud (Syzygium aromaticum) extract used with different extenders on ram spermatozoa during cryopreservation.

    Science.gov (United States)

    Baghshahi, H; Riasi, A; Mahdavi, A H; Shirazi, A

    2014-12-01

    Clove bud (Syzygium aromaticum) extract was added at concentrations of 0, 35, 75, and 115 μg/ml to ovine semen extenders in order to investigate the antioxidant activities of clove bud extract and its effects on semen quality parameters after cryopreservation of ram spermatozoa. The basic extender was composed of Tris, egg yolk, and glycerol. Two other extenders were prepared by substitution of egg yolk with either LDL or egg yolk+SDS. The DPPH inhibition test was employed to assess the antioxidant activity of clove bud extract. Results showed that, compared to vitamin E, clove bud extract had a higher antioxidant activity. Better sperm motility and movement characteristics (Pclove bud extract to semen extenders. Sperm viability and plasma membrane integrity were also higher (Pclove buds extract after cryopreservation processes. Higher levels of clove bud extract, however, had adverse effects on all the sperm quality parameters and significantly reduced (Pclove bud extract had an antioxidant potential that makes it useful for addition to semen extenders and that the best results are obtained with a maximum clove bud extract of 75 μg/ml. Moreover, the combination of egg yolk and a detergent was found to improve sperm quality after the cooling and freeze-thawing processes. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. The adverse prognostic effect of tumor budding on the evolution of cutaneous head and neck squamous cell carcinoma.

    Science.gov (United States)

    Gonzalez-Guerrero, Miriam; Martínez-Camblor, Pablo; Vivanco, Blanca; Fernández-Vega, Ivan; Munguía-Calzada, Pablo; Gonzalez-Gutierrez, Maria Paz; Rodrigo, Juan Pablo; Galache, Cristina; Santos-Juanes, Jorge

    2017-06-01

    Tumor budding is a readily detectable histopathologic feature that has been recognized as an adverse prognostic factor in several human cancers. We sought to assess the correlation of tumor budding with the clinicopathologic features and the prognostic value of tumor budding in cutaneous squamous cell carcinoma (cSCC). Forty-nine primary nonmetastatic and 49 primary metastatic cSCCs to regional lymph nodes were retrospectively studied. Statistical analyses were carried out to assess the relationship between tumor budding, clinicopathologic parameters, and patient survival. Tumor budding was observed in 45 cases of 98 (46%). High-intensity budding (≥5 tumor buds) was observed in 20 tumors. Presence of tumor buds was a significant risk factor for nodal metastasis with crude and adjusted hazard ratios (HRs) of 8.92 (95% CI, 4.39-18.1) and 6.93 (95% CI, 3.30-14.5), respectively, and for reduced overall survival time (crude and adjusted HRs of 2.03 [95% CI, 1.26-3.28] and 1.72 [95% CI, 1.05-2.83], respectively). This was a retrospective study limited to cSCCs of the head and neck. Examined tumors were >2 mm thick, and all were from a primary excision. These results indicate an increased frequency of nodal metastasis and risk of death in patients with tumor buds. Copyright © 2017 American Academy of Dermatology, Inc. Published by Elsevier Inc. All rights reserved.

  10. Distinct roles of Hand2 in initiating polarity and posterior Shh expression during the onset of mouse limb bud development.

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    Antonella Galli

    2010-04-01

    Full Text Available The polarization of nascent embryonic fields and the endowment of cells with organizer properties are key to initiation of vertebrate organogenesis. One such event is antero-posterior (AP polarization of early limb buds and activation of morphogenetic Sonic Hedgehog (SHH signaling in the posterior mesenchyme, which in turn promotes outgrowth and specifies the pentadactylous autopod. Inactivation of the Hand2 transcriptional regulator from the onset of mouse forelimb bud development disrupts establishment of posterior identity and Shh expression, which results in a skeletal phenotype identical to Shh deficient limb buds. In wild-type limb buds, Hand2 is part of the protein complexes containing Hoxd13, another essential regulator of Shh activation in limb buds. Chromatin immunoprecipitation shows that Hand2-containing chromatin complexes are bound to the far upstream cis-regulatory region (ZRS, which is specifically required for Shh expression in the limb bud. Cell-biochemical studies indicate that Hand2 and Hoxd13 can efficiently transactivate gene expression via the ZRS, while the Gli3 repressor isoform interferes with this positive transcriptional regulation. Indeed, analysis of mouse forelimb buds lacking both Hand2 and Gli3 reveals the complete absence of antero-posterior (AP polarity along the entire proximo-distal axis and extreme digit polydactyly without AP identities. Our study uncovers essential components of the transcriptional machinery and key interactions that set-up limb bud asymmetry upstream of establishing the SHH signaling limb bud organizer.

  11. Reactivation from latency displays HIV particle budding at plasma membrane, accompanying CD44 upregulation and recruitment

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    Sano Kouichi

    2009-07-01

    Full Text Available Abstract Background It has been accepted that HIV buds from the cell surface in T lymphocytes, whereas in macrophages it buds into intracellular endosomes. Recent studies, on the other hand, suggest that HIV preferentially buds from the cell surface even in monocytic cells. However, most studies are based on observations in acutely infected cells and little is known about HIV budding concomitant with reactivation from latency. Such studies would provide a better understanding of a reservoir for HIV. Results We observed HIV budding in latently infected T lymphocytic and monocytic cell lines following TNF-α stimulation and examined the upregulation of host factors that may be involved in particle production. Electron microscopy analysis revealed that reactivation of latently infected J1.1 cells (latently infected Jurkat cells with HIV-1 and U1 cells (latently infected U937 cells with HIV-1 displayed HIV particle budding predominantly at the plasma membrane, a morphology that is similar to particle budding in acutely infected Jurkat and U937 cells. When mRNA expression levels were quantified by qRT-PCR, we found that particle production from reactivated J1.1 and U1 cells was accompanied by CD44 upregulation. This upregulation was similarly observed when Jurkat and U937 cells were acutely infected with HIV-1 but not when just stimulated with TNF-α, suggesting that CD44 upregulation was linked with HIV production but not with cell stimulation. The molecules in endocytic pathways such as CD63 and HRS were also upregulated when U1 cells were reactivated and U937 cells were acutely infected with HIV-1. Confocal microscopy revealed that these upregulated host molecules were recruited to and accumulated at the sites where mature particles were formed at the plasma membrane. Conclusion Our study indicates that HIV particles are budded at the plasma membrane upon reactivation from latency, a morphology that is similar to particle budding in acute

  12. A2BR adenosine receptor modulates sweet taste in circumvallate taste buds.

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    Shinji Kataoka

    Full Text Available In response to taste stimulation, taste buds release ATP, which activates ionotropic ATP receptors (P2X2/P2X3 on taste nerves as well as metabotropic (P2Y purinergic receptors on taste bud cells. The action of the extracellular ATP is terminated by ectonucleotidases, ultimately generating adenosine, which itself can activate one or more G-protein coupled adenosine receptors: A1, A2A, A2B, and A3. Here we investigated the expression of adenosine receptors in mouse taste buds at both the nucleotide and protein expression levels. Of the adenosine receptors, only A2B receptor (A2BR is expressed specifically in taste epithelia. Further, A2BR is expressed abundantly only in a subset of taste bud cells of posterior (circumvallate, foliate, but not anterior (fungiform, palate taste fields in mice. Analysis of double-labeled tissue indicates that A2BR occurs on Type II taste bud cells that also express Gα14, which is present only in sweet-sensitive taste cells of the foliate and circumvallate papillae. Glossopharyngeal nerve recordings from A2BR knockout mice show significantly reduced responses to both sucrose and synthetic sweeteners, but normal responses to tastants representing other qualities. Thus, our study identified a novel regulator of sweet taste, the A2BR, which functions to potentiate sweet responses in posterior lingual taste fields.

  13. A2BR Adenosine Receptor Modulates Sweet Taste in Circumvallate Taste Buds

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    Yang, Dan; Shultz, Nicole; Vandenbeuch, Aurelie; Ravid, Katya; Kinnamon, Sue C.; Finger, Thomas E.

    2012-01-01

    In response to taste stimulation, taste buds release ATP, which activates ionotropic ATP receptors (P2X2/P2X3) on taste nerves as well as metabotropic (P2Y) purinergic receptors on taste bud cells. The action of the extracellular ATP is terminated by ectonucleotidases, ultimately generating adenosine, which itself can activate one or more G-protein coupled adenosine receptors: A1, A2A, A2B, and A3. Here we investigated the expression of adenosine receptors in mouse taste buds at both the nucleotide and protein expression levels. Of the adenosine receptors, only A2B receptor (A2BR) is expressed specifically in taste epithelia. Further, A2BR is expressed abundantly only in a subset of taste bud cells of posterior (circumvallate, foliate), but not anterior (fungiform, palate) taste fields in mice. Analysis of double-labeled tissue indicates that A2BR occurs on Type II taste bud cells that also express Gα14, which is present only in sweet-sensitive taste cells of the foliate and circumvallate papillae. Glossopharyngeal nerve recordings from A2BR knockout mice show significantly reduced responses to both sucrose and synthetic sweeteners, but normal responses to tastants representing other qualities. Thus, our study identified a novel regulator of sweet taste, the A2BR, which functions to potentiate sweet responses in posterior lingual taste fields. PMID:22253866

  14. Androgenic regulation of ventral epithelial bud number and pattern in mouse urogenital sinus.

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    Allgeier, Sarah H; Lin, Tien-Min; Moore, Robert W; Vezina, Chad M; Abler, Lisa L; Peterson, Richard E

    2010-02-01

    The ventral urogenital sinus (UGS) of control male mice has two rows of 3-4 prostatic buds at birth, but how androgens regulate ventral bud (VB) number and patterning is unclear. VBs in both sexes appeared to be a mixture of prostatic and urethral buds. UGSs from Tfm male and antiandrogen (flutamide)-exposed mice had small VBs, suggesting that initiation of some VBs is androgen independent. Tfm male mice are widely considered completely androgen insensitive yet their UGSs were 5alpha-dihydrotestosterone (DHT)- responsive. VBs (6-8) were generally distributed bimodally on the left-right axis at both minimal and normal male androgen signaling. Yet control females and DHT-exposed Tfm males had 13-14 VBs, whose left-right distribution was fairly uniform. These results suggest that VB number and distribution respond biphasically as androgen signaling increases from minimal, and that androgens regulate bud specification. Complete VB agenesis by the selective budding inhibitor 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) required high androgen signaling.

  15. Ulex Europaeus Agglutinin-1 Is a Reliable Taste Bud Marker for In Situ Hybridization Analyses.

    Science.gov (United States)

    Yoshimoto, Joto; Okada, Shinji; Kishi, Mikiya; Misaka, Takumi

    2016-03-01

    Taste signals are received by taste buds. To better understand the taste reception system, expression patterns of taste-related molecules are determined by in situ hybridization (ISH) analyses at the histological level. Nevertheless, even though ISH is essential for determining mRNA expression, few taste bud markers can be applied together with ISH. Ulex europaeus agglutinin-1 (UEA-1) appears to be a reliable murine taste bud marker based on immunohistochemistry (IHC) analyses. However, there is no evidence as to whether UEA-1 can be used for ISH. Thus, the present study evaluated UEA-1 using various histochemical methods, especially ISH. When lectin staining was performed after ISH procedures, UEA-1 clearly labeled taste cellular membranes and distinctly indicated boundaries between taste buds and the surrounding epithelial cells. Additionally, UEA-1 was determined as a taste bud marker not only when used in single-colored ISH but also when employed with double-labeled ISH or during simultaneous detection using IHC and ISH methods. These results suggest that UEA-1 is a useful marker when conducting analyses based on ISH methods. To clarify UEA-1 staining details, multi-fluorescent IHC (together with UEA-1 staining) was examined, resulting in more than 99% of cells being labeled by UEA-1 and overlapping with KCNQ1-expressing cells. © 2016 The Histochemical Society.

  16. Effect of the cytokinins combination on adventitious buds formation in banana cv. 'Gros Michel' (Musa AAA

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    Idalmis Bermúdez-Caraballoso

    2017-03-01

    Full Text Available Cytokinins are used for adventitious buds formation but high concentrations may have negative effects on plant regeneration. The aim of the present investigation was to determine the effect of different combinations of cytokinins on the induction of adventitious buds of cultivar ‘Gros Michel’ (Musa AAA. Different concentrations of 6-Benzylaminopurine (2.0, 4.0 and 6.0 mg l-1 and Thidiazuron (0.6, 0.8, 1.0 mg l-1 were studied to achieve the formation of these structures. As controls, 22.5 mg l-1 6-BAP and 2.0 mg l-1 TDZ, widely used for these purposes in this culture, were used. The treatment with 2.0 mg l-1 6-BAP + 1.0 mg l-1 TDZ was the one that achieved the highest percentage of adventitious buds formation, with 80% of the explants, with results much higher than those achieved by the controls and other combinations studied. With this treatment, an average of 2.72 adventitious buds per explant was obtained with the least number of shoots per explant (1.24 at 60 days of culture. For this reason, this combination of cytokinins was selected for the formation of adventitious buds in banana cv. 'Gros Michel', as it was possible to decrease the levels of 6-BAP and TDZ that have negative effects on the subsequent plants regeneration.   Keywords: bananas, 6- Benzylaminopurine, Thidiazuron

  17. Overexpression of DEMETER, a DNA demethylase, promotes early apical bud maturation in poplar.

    Science.gov (United States)

    Conde, Daniel; Moreno-Cortés, Alicia; Dervinis, Christopher; Ramos-Sánchez, José M; Kirst, Matias; Perales, Mariano; González-Melendi, Pablo; Allona, Isabel

    2017-11-01

    The transition from active growth to dormancy is critical for the survival of perennial plants. We identified a DEMETER-like (CsDML) cDNA from a winter-enriched cDNA subtractive library in chestnut (Castanea sativa Mill.), an economically and ecologically important species. Next, we characterized this DNA demethylase and its putative ortholog in the more experimentally tractable hybrid poplar (Populus tremula × alba), under the signals that trigger bud dormancy in trees. We performed phylogenetic and protein sequence analysis, gene expression profiling, and 5-methyl-cytosine methylation immunodetection studies to evaluate the role of CsDML and its homolog in poplar, PtaDML6. Transgenic hybrid poplars overexpressing CsDML were produced and analysed. Short days and cold temperatures induced CsDML and PtaDML6. Overexpression of CsDML accelerated short-day-induced bud formation, specifically from Stages 1 to 0. Buds acquired a red-brown coloration earlier than wild-type plants, alongside with the up-regulation of flavonoid biosynthesis enzymes and accumulation of flavonoids in the shoot apical meristem and bud scales. Our data show that the CsDML gene induces bud formation needed for the survival of the apical meristem under the harsh conditions of winter. © 2017 John Wiley & Sons Ltd.

  18. Tumor Budding, uPA, and PAI-1 in Colorectal Cancer: Update of a Prospective Study

    Science.gov (United States)

    Hardt, Jochen; Franz, Simon; Schaller, Tina; Schenkirsch, Gerhard; Kriening, Bernadette; Hoffmann, Reinhard; Rüth, Stefan

    2017-01-01

    Aims. The prognostic role of the proteases uPA and PAI-1, as well as tumor budding, in colon cancer, has been investigated previously. Methods. We provide 6-year follow-up data and results of the validation set. The initial test set and validation set consisted of 55 colon cancers and 68 colorectal cancers, respectively. Tissue samples were analyzed for uPA and PAI-1 using a commercially available Enzyme-Linked Immunosorbent Assay (ELISA). Tumor budding was analyzed on cytokeratin-stained slides. Survival analyses were performed using cut-offs that were determined previously. Results. uPA was not prognostic for outcome. PAI-1 showed a trend towards reduced cancer specific survival in PAI-1 high-grade cases (68 versus 83 months; P = 0.091). The combination of high-grade PAI-1 and tumor budding was associated with significantly reduced cancer specific survival (60 versus 83 months; P = 0.021). After pooling the data from both sets, multivariate analyses revealed that the factors pN-stage, V-stage, and a combination of tumor budding and PAI-1 were independently prognostic for the association with distant metastases. Conclusions. A synergistic adverse effect of PAI-1 and tumor budding in uni- and multivariable analyses was found. PAI-1 could serve as a target for anticancer therapy. PMID:28286517

  19. Tumor Budding, uPA, and PAI-1 in Colorectal Cancer: Update of a Prospective Study

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    Bruno Märkl

    2017-01-01

    Full Text Available Aims. The prognostic role of the proteases uPA and PAI-1, as well as tumor budding, in colon cancer, has been investigated previously. Methods. We provide 6-year follow-up data and results of the validation set. The initial test set and validation set consisted of 55 colon cancers and 68 colorectal cancers, respectively. Tissue samples were analyzed for uPA and PAI-1 using a commercially available Enzyme-Linked Immunosorbent Assay (ELISA. Tumor budding was analyzed on cytokeratin-stained slides. Survival analyses were performed using cut-offs that were determined previously. Results. uPA was not prognostic for outcome. PAI-1 showed a trend towards reduced cancer specific survival in PAI-1 high-grade cases (68 versus 83 months; P=0.091. The combination of high-grade PAI-1 and tumor budding was associated with significantly reduced cancer specific survival (60 versus 83 months; P=0.021. After pooling the data from both sets, multivariate analyses revealed that the factors pN-stage, V-stage, and a combination of tumor budding and PAI-1 were independently prognostic for the association with distant metastases. Conclusions. A synergistic adverse effect of PAI-1 and tumor budding in uni- and multivariable analyses was found. PAI-1 could serve as a target for anticancer therapy.

  20. Progression level of extracapsular spread and tumor budding for cervical lymph node metastasis of OSCC.

    Science.gov (United States)

    Yamada, Shin-Ichi; Otsuru, Mitsunobu; Yanamoto, Souichi; Hasegawa, Takumi; Aizawa, Hitoshi; Kamata, Takahiro; Yamakawa, Nobuhiro; Kohgo, Tomoyuki; Ito, Akira; Noda, Yuri; Hirai, Chihoko; Kitamura, Tetsuya; Okura, Masaya; Kirita, Tadaaki; Ueda, Michihiro; Yamashita, Tetsuro; Ota, Yoshihide; Komori, Takahide; Umeda, Masahiro; Kurita, Hiroshi

    2017-10-06

    The progression level of extracapsular spread (ECS) for cervical lymph node metastasis of oral squamous cell carcinoma (OSCC) was previously divided into three types, and their relationships with the prognosis of patients were re-examined. The Kaplan-Meier method was used to examine overall survival (OS) and relapse-free survival (RFS) curves. Prognosis factor for recurrence was analyzed with univariate and multivariate analysis. ECS was detected in 216 cases of OSCC and analyzed. The 5-year overall survival and RFS rates of patients with type C, which was microscopically defined as tumor invasion to perinodal fat or muscle tissue, were significantly poor at 40.6 and 37.8%, respectively. The results of a univariate analysis suggested that the prognosis of ECS in OSCC patients is associated with its progression level, particularly type C. The 5-year RFS rate of type C with tumor budding was significantly poor at 31.5%. Type C with tumor budding correlated with local and regional recurrence as well as distant metastasis. In a multivariate analysis, tumor budding was identified as an independent prognostic factor. These results suggest that the progression level of ECS and tumor budding are useful prognostic factors in OSCC patients. This study indicated that the progression level and tumor budding of ECS for cervical lymph node metastasis were useful prognostic factors in OSCC patients.

  1. Histopathological Significance and Prognostic Impact of Tumor Budding in Colorectal Cancer.

    Science.gov (United States)

    Mehta, Anurag; Goswami, Malini; Sinha, Rupal

    2017-03-01

    Colorectal cancer (CRC) is a heterogeneous disease with complex etiology. New prognostic factors for this group of disease need to be investigated. This study evaluated the histopathological significance and prognostic impact of tumor budding in CRC. A total of 60 treatment naive consecutive patients undergoing surgical resection for CRC during the period of January 2011 to December 2013 were included in the study. Details of each patient related to their demographic and tumor profile were recorded. Pan Cytokeratin immunohistochemistry was applied on chosen sections and tumor budding was assessed. The most frequent site of involvement was rectosigmoid and sigmoid colon (31.6%). The majority of the cases were moderately differentiated (75%) in morphology, and showed tumor invasion into the pericolic/subserosal fat (66.6%), and were stage III (38.3%). Nodal involvement was present in 50% cases. Correlations between tumor budding and nodal involvement (p-value 0.039) and AJCC stage (p-value 0.021) were found to be statistically significant. Tumor budding may be a promising and powerful predictor of lymphnodal metastasis and a higher stage of tumor and can be used as a marker for assessing the aggressiveness of CRC. Routine hemotoxylin-eosin staining supported by cytokeratin immunostain can aid in the grading of tumor budding in CRC. © 2017 by the Association of Clinical Scientists, Inc.

  2. Bud break responds more strongly to daytime than night-time temperature under asymmetric experimental warming.

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    Rossi, Sergio; Isabel, Nathalie

    2017-01-01

    Global warming is diurnally asymmetric, leading to a less cold, rather than warmer, climate. We investigated the effects of asymmetric experimental warming on plant phenology by testing the hypothesis that daytime warming is more effective in advancing bud break than night-time warming. Bud break was monitored daily in Picea mariana seedlings belonging to 20 provenances from Eastern Canada and subjected to daytime and night-time warming in growth chambers at temperatures varying between 8 and 16 °C. The higher advancements of bud break and shorter times required to complete the phenological phases occurred with daytime warming. Seedlings responded to night-time warming, but still with less advancement of bud break than under daytime warming. No advancement was observed when night-time warming was associated with a daytime cooling. The effect of the treatments was uniform across provenances. Our observations realized under controlled conditions allowed to experimentally demonstrate that bud break can advance under night-time warming, but to a lesser extent than under daytime warming. Prediction models using daily timescales could neglect the diverging influence of asymmetric warming and should be recalibrated for higher temporal resolutions. © 2016 John Wiley & Sons Ltd.

  3. The bud break process and its variation among local populations of boreal black spruce.

    Science.gov (United States)

    Rossi, Sergio; Bousquet, Jean

    2014-01-01

    Phenology of local populations can exhibit adaptations to the current environmental conditions resulting from a close interaction between climate and genotype. The bud break process and its variations among populations were analyzed in greenhouse by monitoring the growth resumption in black spruce [Picea mariana (Mill.) BSP] seedlings originating from seeds of five stands across the closed boreal forest in Quebec, Canada. Bud break lasted 15 days and occurred earlier and quicker in northern provenances. Provenance explained between 10.2 and 32.3% of the variance in bud break, while the families accounted for a smaller but still significant part of the variance. The late occurrence of one phenological phase corresponded to a delayed occurrence of the others according to linear relationships. A causal model was proposed in the form of a chain of events with each phase of bud break being related to the previous and successive one, while no link was observed between non-adjacent phases. The adaptation of black spruce populations along the latitudinal gradient points toward a strategy based on rapid physiological processes triggered by temperature increase inducing high metabolic activity. The variation observed in bud break reflects an evolutionary trade-off between maximization of security and taking advantage of the short growing season. This work provides evidence of the phenological adaptations of black spruce to its local environmental conditions while retaining sizeable genetic diversity within populations. Because of the multigenic nature of phenology, this diversity should provide some raw material for adaptation to changing local environmental conditions.

  4. Galectin-3 promotes HIV-1 budding via association with Alix and Gag p6

    Science.gov (United States)

    Wang, Sheng-Fan; Tsao, Ching-Han; Lin, Yu-Ting; Hsu, Daniel K; Chiang, Meng-Lin; Lo, Chia-Hui; Chien, Fan-Ching; Chen, Peilin; Arthur Chen, Yi-Ming; Chen, Huan-Yuan; Liu, Fu-Tong

    2014-01-01

    Galectin-3 has been reported to regulate the functions of a number of immune cell types. We previously reported that galectin-3 is translocated to immunological synapses in T cells upon T-cell receptor engagement, where it associates with ALG-2-interacting protein X (Alix). Alix is known to coordinate with the endosomal sorting complex required for transport (ESCRT) to promote human immunodeficiency virus (HIV)-1 virion release. We hypothesized that galectin-3 plays a role in HIV-1 viral budding. Cotransfection of cells of the Jurkat T line with galectin-3 and HIV-1 plasmids resulted in increased HIV-1 budding, and suppression of galectin-3 expression by RNAi in Hut78 and primary CD4+ T cells led to reduced HIV-1 budding. We used immunofluorescence microscopy to observe the partial colocalization of galectin-3, Alix and Gag in HIV-1-infected cells. Results from co-immunoprecipitation experiments indicate that galectin-3 expression promotes Alix-Gag p6 association, whereas the results of Alix knockdown suggest that galectin-3 promotes HIV-1 budding through Alix. HIV-1 particles released from galectin-3-expressing cells acquire the galectin-3 protein in an Alix-dependent manner, with proteins primarily residing inside the virions. We also found that the galectin-3 N-terminal domain interacts with the proline-rich region of Alix. Collectively, these results suggest that endogenous galectin-3 facilitates HIV-1 budding by promoting the Alix-Gag p6 association. PMID:24996823

  5. Transcriptome and Metabolite Changes during Hydrogen Cyanamide-Induced Floral Bud Break in Sweet Cherry

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    Irina A. Ionescu

    2017-07-01

    Full Text Available Release of bud dormancy in perennial woody plants is a temperature-dependent process and thus flowering in these species is heavily affected by climate change. The lack of cold winters in temperate growing regions often results in reduced flowering and low fruit yields. This is likely to decrease the availability of fruits and nuts of the Prunus spp. in the near future. In order to maintain high yields, it is crucial to gain detailed knowledge on the molecular mechanisms controlling the release of bud dormancy. Here, we studied these mechanisms using sweet cherry (Prunus avium L., a crop where the agrochemical hydrogen cyanamide (HC is routinely used to compensate for the lack of cold winter temperatures and to induce flower opening. In this work, dormant flower buds were sprayed with hydrogen cyanamide followed by deep RNA sequencing, identifying three main expression patterns in response to HC. These transcript level results were validated by quantitative real time polymerase chain reaction and supported further by phytohormone profiling (ABA, SA, IAA, CK, ethylene, JA. Using these approaches, we identified the most up-regulated pathways: the cytokinin pathway, as well as the jasmonate and the hydrogen cyanide pathway. Our results strongly suggest an inductive effect of these metabolites in bud dormancy release and provide a stepping stone for the characterization of key genes in bud dormancy release.

  6. Cell polarity in Saccharomyces cerevisiae depends on proper localization of the Bud9 landmark protein by the EKC/KEOPS complex.

    Science.gov (United States)

    Kato, Yu; Kawasaki, Hiroshi; Ohyama, Yoshifumi; Morishita, Takashi; Iwasaki, Hiroshi; Kokubo, Tetsuro; Hirano, Hisashi

    2011-08-01

    In diploid Saccharomyces cerevisiae cells, bud-site selection is determined by two cortical landmarks, Bud8p and Bud9p, at the distal and proximal poles, respectively. Their localizations depend on the multigenerational proteins Rax1p/Rax2p. Many genes involved in bud-site selection were identified previously by genome-wide screening of deletion mutants, which identified BUD32 that causes a random budding in diploid cells. Bud32p is an atypical kinase involved in a signaling cascade of Sch9p kinase, the yeast homolog of Akt/PKB, and a component of the EKC/KEOPS (endopeptidase-like, kinase, chromatin-associated/kinase, putative endopeptidase, and other proteins of small size) complex that functions in telomere maintenance and transcriptional regulation. However, its role in bipolar budding has remained unclear. In this report, we show that the Sch9p kinase cascade does not affect bipolar budding but that the EKC/KEOPS complex regulates the localization of Bud9p. The kinase activity of Bud32p, which is essential for the functions of the EKC/KEOPS complex but is not necessary for the Sch9p signaling cascade, is required for bipolar bud-site selection. BUD9 is necessary for random budding in each deletion mutant of EKC/KEOPS components, and RAX2 is genetically upstream of EKC/KEOPS genes for the regulation of bipolar budding. The asymmetric localization of Bud9p was dependent on the complex, but Bud8p and Rax2p were not. We concluded that the EKC/KEOPS complex is specifically involved in the regulation of Bud9p localization downstream of Rax1p/Rax2p.

  7. Ferrite-Cored Solenoidal Induction Coil Sensor for BUD (MM-1667)

    Energy Technology Data Exchange (ETDEWEB)

    Morrison, F.; Becker, A.; Conti, U.; Gasperikova, E.

    2011-06-15

    We have designed and lab tested a new ferrite cored induction coil sensor for measuring the secondary fields from metallic UXO with the BUD system. The objective was to replace the 5-inch diameter air-cored coils in the BUD system with smaller sensors that would allow the placement of multiple sensors in the smaller package of the new BUD hand-held system. A ferrite-cored solenoidal coil of length L can easily be made to have sensitivity and noise level roughly the same as an air-cored coil of a diameter on the same order as L. A ferrite-cored solenoidal coil can easily have a feedback configuration to achieve critical damping. The feedback configuration leads to a very stable response. Feedback ferrite-cored solenoidal coils show very little interaction as long as they are separated by one half their length.

  8. The bud rot of oil palm in San Lorenzo, Esmeraldas province, Ecuador

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    Fernando Rivas Figueroa

    2015-01-01

    Full Text Available Oil palms (Elaeis guineensis Jacq in the area of San Lorenzo were directly observed, and some plants were dissected to assess the internal symptoms, with the purpose of characterizing the symptomatology of bud rot. The plants showed chlorosis and yellowing of young leaves around the bud, necrosis of leaflets in young leaves, necrosis and rot of spears (outer leaf in the process of opening, bending of spears leaves due to the breaking in the lower third, necrosis and internal bud decay which can descend to the meristem and leads to the death of the plants. It is estimated that the incidence of the disease ranges around 30 %. In Ecuador, Researchers have not still identified the agent causing this disease since the early 80’s.

  9. High autumn temperature delays spring bud burst in boreal trees, counterbalancing the effect of climatic warming

    Energy Technology Data Exchange (ETDEWEB)

    Heide, O. M. [Agricultural Univesity of Norway, Department of Biology and Nature Conservation, As (Norway)

    2003-09-01

    The effect of temperature during short-day dormancy induction on the duration and stability of bud dormancy was examined in three boreal tree species (2 birches and 1 alder) grown in a controlled environment. The phenology of the latitudinal range of birch populations, and the relationship between spring bud burst and autumn and spring temperatures were also studied. Results showed that during short-day dormancy induction in the autumn high temperatures delayed bud burst in the following spring in both controlled and natural environments. It is suggested that this response to higher autumn temperatures may be a manifestation of a general synergism between high temperature and short-day photoperiodic processes, and may be an adaptive mechanism common to boreal trees. It is further conjectured that this mechanism may be important in counterbalancing the potentially adverse effects of higher winter temperatures on dormancy stability of boreal trees during climate warming. 23 refs., 2 tabs., 4 figs.

  10. Progress and renewal in gustation: new insights into taste bud development.

    Science.gov (United States)

    Barlow, Linda A

    2015-11-01

    The sense of taste, or gustation, is mediated by taste buds, which are housed in specialized taste papillae found in a stereotyped pattern on the surface of the tongue. Each bud, regardless of its location, is a collection of ∼100 cells that belong to at least five different functional classes, which transduce sweet, bitter, salt, sour and umami (the taste of glutamate) signals. Taste receptor cells harbor functional similarities to neurons but, like epithelial cells, are rapidly and continuously renewed throughout adult life. Here, I review recent advances in our understanding of how the pattern of taste buds is established in embryos and discuss the cellular and molecular mechanisms governing taste cell turnover. I also highlight how these findings aid our understanding of how and why many cancer therapies result in taste dysfunction. © 2015. Published by The Company of Biologists Ltd.

  11. Greenhouse irrigation water depths in relation to rose stem and bud qualities

    Directory of Open Access Journals (Sweden)

    Folegatti Marcos Vinícius

    2001-01-01

    Full Text Available The cultivation of roses occupies a special place in the flower production of Brazil, the concern with the quality of the buds being intimately related with the appropriate supply of water and nutrients to the plant. With the objective of evaluating stem and bud quality the rose variety 'Osiana' was cultivated in a greenhouse using different irrigation water depths based on fractions of pan evaporation (0.25, 0.50, 0.75, 1.00 and 1.25. The experimental design consisted of total randomized blocks with five replications and five treatments. There is a linear tendency of increasing the length and diameter of the stems and the length and diameter of the buds with increasing irrigation water depths.

  12. Direct cryopreservation of winter buds of nine cultivars of blackcurrant (Ribes nigrum L).

    Science.gov (United States)

    Green, Jon; Grout, Brian

    2010-01-01

    Direct cryopreservation of overwintering, dormant buds has been applied to nine blackcurrant cultivars, using a 7 day dehydration period at c.-20° C before plunging directly into liquid nitrogen. The buds on shoots thawed from -20° C showed normal development simply by standing them in water and all the cultivars could be successfully recovered (> 58%) by grafting. None of the shoots thawed from liquid nitrogen showed any development after standing in water and all of the grafts failed. Shoots thawed from liquid nitrogen showed significant damage to xylem transport, and the cortical tissues necessary fro successful grafting showed significant loss of membrane semipermeability. However, buds excised from shoots immediately after thawing from liquid nitrogen were viable and could be recovered using in vitro culture. Survival ranged from 88 to 55%, depending upon cultivar.

  13. Multiple Enhancers Regulate Hoxd Genes and the Hotdog LncRNA during Cecum Budding

    Directory of Open Access Journals (Sweden)

    Saskia Delpretti

    2013-10-01

    Full Text Available Hox genes are required for the development of the intestinal cecum, a major organ of plant-eating species. We have analyzed the transcriptional regulation of Hoxd genes in cecal buds and show that they are controlled by a series of enhancers located in a gene desert flanking the HoxD cluster. The start site of two opposite long noncoding RNAs (lncRNAs, Hotdog and Twin of Hotdog, selectively contacts the expressed Hoxd genes in the framework of a topological domain, coinciding with robust transcription of these genes during cecum budding. Both lncRNAs are specifically transcribed in the cecum, albeit bearing no detectable function in trans. Hedgehogs have kept this regulatory potential despite the absence of the cecum, suggesting that these mechanisms are used in other developmental situations. In this context, we discuss the implementation of a common “budding toolkit” between the cecum and the limbs.

  14. ESCRT-independent budding of HIV-1 gag virus-like particles from Saccharomyces cerevisiae spheroplasts.

    Directory of Open Access Journals (Sweden)

    Andrew P Norgan

    Full Text Available Heterologous expression of HIV-1 Gag in a variety of host cells results in its packaging into virus-like particles (VLPs that are subsequently released into the extracellular milieu. This phenomenon represents a useful tool for probing cellular factors required for viral budding and has contributed to the discovery of roles for ubiquitin ligases and the endosomal sorting complexes required for transport (ESCRTs in viral budding. These factors are highly conserved throughout eukaryotes and have been studied extensively in the yeast Saccharomyces cerevisiae, a model eukaryote previously utilized as a host for the production of VLPs. We used heterologous expression of HIV Gag in yeast spheroplasts to examine the role of ESCRTs and associated factors (Rsp5, a HECT ubiquitin ligase of the Nedd4 family; Bro1, a homolog of Alix; and Vps4, the AAA-ATPase required for ESCRT function in all contexts/organisms investigated in the generation of VLPs. Our data reveal: 1 characterized Gag-ESCRT interaction motifs (late domains are not required for VLP budding, 2 loss of function alleles of the essential HECT ubiquitin ligase Rsp5 do not display defects in VLP formation, and 3 ESCRT function is not required for VLP formation from spheroplasts. These results suggest that the egress of HIV Gag from yeast cells is distinct from the most commonly described mode of exit from mammalian cells, instead mimicking ESCRT-independent VLP formation observed in a subset of mammalian cells. As such, budding of Gag from yeast cells appears to represent ESCRT-independent budding relevant to viral replication in at least some situations. Thus the myriad of genetic and biochemical tools available in the yeast system may be of utility in the study of this aspect of viral budding.

  15. Colorectal carcinoma tumour budding and podia formation in the xenograft microenvironment.

    Science.gov (United States)

    Prall, Friedrich; Maletzki, Claudia; Hühns, Maja; Krohn, Mathias; Linnebacher, Michael

    2017-01-01

    Tumour budding and podia formation are well-appreciated in surgical pathology as an aggressive invasion phenotype of colorectal carcinoma cells that is attained in the microenvironment of the invasive margin. In this study, we addressed how tumour budding and podia formation feature in xenografts. Primary colorectal carcinomas (N = 44) of various molecular types (sporadic standard type, high-degree microsatellite-unstable, CpG island methylator phenotype) were transplanted subcutaneously into T and B cell-deficient NSG mice, making possible immunohistochemistry with routine surgical pathology antibodies. Tumor budding and podia formation were both appreciably present in the xenografts. Quantitative evaluations of cytokeratin immunostains of primaries and their corresponding xenografts showed a reduction of tumour buds in the xenografts. Furthermore, in xenografts tumour cells were completely negative by pSTAT3 immunohistochemistry, indicating absence of cytokine/chemokine signalling, but nuclear β-catenin and SMAD4 immunostainings as read-out of wnt and BMP pathway activation, respectively, were maintained. Carcinoma cells in most xenografts retained immunostaining of at least some nuclei by immunohistochemistry with antibodies against pERK1/2. K-ras/B-raf mutational status did not correlate with tumour budding or podia formation in the xenografts. Our results indicate that tumour budding and podia formation can be modelled by xenografting, and in NSG mice it can be studied with the same immunohistochemical methods as used for primaries in surgical pathology. Dysregulation of wnt and BMP signalling appears to be transferred into the xenograft microenvironment, but not cytokine/chemokine signalling.

  16. Database Description - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available List Contact us Budding yeast cDNA sequencing project Database Description General information of database D...ases Organism Taxonomy Name: Saccharomyces cerevisiae Taxonomy ID: 4932 Database description 5'-end sequence...nuine 5'-end, mapping the 5'-end sequence to the genome will lead to accurate identification of the transcript... title: A large-scale full-length cDNA analysis to explore the budding yeast transcriptome. Author name(s): ...rvices Not available URL of Web services - Need for user registration - About This Database Database Descript

  17. Download - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available English ]; } else { document.getElementById(lang).innerHTML= '[ Japanese | English ]'; } } window.onload = ...List Contact us Budding yeast cDNA sequencing project Download First of all, please read the license of this... database. Data names and data descriptions are about the downloadable data in this page. They might not cor... # Data name File Simple search and download 1 README README_e.html - 2 5'-end se...quences of budding yeast full-length cDNA clones and quality scores yeast_seq_qual.zip (59.9MB) Simple search and dow

  18. CO2 mediated interaction in yeast stimulates budding and growth on minimal media.

    Directory of Open Access Journals (Sweden)

    Ilya V Volodyaev

    Full Text Available Here we show that carbon dioxide (CO2 stimulates budding and shortens the lag-period of Saccharomyces cerevisiae cultures, grown on specific weak media. CO2 can be both exogenous and secreted by another growing yeast culture. We also show that this effect can be observed only in the lag-period, and demonstrate minimal doses and duration of culture exposition to CO2. Opposite to the effects of CO2 sensitivity, previously shown for pathogens, where increased concentration of CO2 suppressed mitosis and stimulated cell differentiation and invasion, here it stimulates budding and culture growth.

  19. Assessment of tumor budding in colorectal carcinoma: correlation with β-catenin nuclear expression.

    Science.gov (United States)

    El-Gendi, S; Al-Gendi, A

    2011-03-01

    Tumor budding (TB) is showing increasing promise as a colorectal cancer (CRC) prognosticator that is independent of TNM staging. β-Catenin is a component of the Wingless/Wnt signaling pathway that is bound to membrane-associated E-cadherin and is essential for its correct position and function. This study was designed to detect TB in 44 resected primary CRC cases and also to compare β-catenin expression in the tumor budding sites (TBS) and in the tumor center. Tumor budding was assessed in both H&E and pankeratin immunostained sections. Agreement between TB scoring using pancytokeratin and H&E was tested. Also, typing of the tumor margin and determination of degree of cytoplasmic pseudo-fragmentation was done. Tumor budding, cytoplasmic pseudofragments and β-catenin expression were related to known CRC prognosticators. Ten tumors (22.7%) showed low grade (LG) budding and 34 tumors (77.3%) showed high grade (HG) budding. The 34 HG budding tumors were further subdivided into moderate and severe (n=13, n=21, respectively) budding cancers. Twenty nine tumors (65.9%) showed LG cytoplasmic pseudofragments and 15 tumors (34.1%) showed HG pseudofragments. Scoring of TB on H&E and pankeratin stained sections revealed moderate agreement (Kappa=.558; p=tumor center (p=.005, p=.000, respectively). In opposition, membranous β-catenin expression was significantly higher in the tumor center than in TBS (p=.001). Although, nuclear β-catenin accumulation at TBS was noted, yet, it did not relate significantly with both TB and cytoplasmic pseudofragments around TBS (p=.649; p=.675, respectively). Also, nuclear β-catenin immunoreactivity did not relate significantly with the various clinicopathological variables. Pankeratin immunostaining facilitates typing of CRC invasive margin, and determination of the degree of TB and cytoplasmic pseudo-fragmentation. β-Catenin expression differs significantly between tumor center and TBS in CRC. Cut-offs for TB assessment should be unified

  20. Safety assessment of a standardized polyphenolic extract of clove buds: Subchronic toxicity and mutagenicity studies

    OpenAIRE

    Vijayasteltar, Liju; Nair, Gopakumar Gopinathan; Maliakel, Balu; Kuttan, Ramadasan; I.M., Krishnakumar

    2016-01-01

    Despite the various reports on the toxicity of clove oil and its major component eugenol, systematic evaluations on the safety of polyphenolic extracts of clove buds have not been reported. Considering the health beneficial pharmacological effects and recent use of clove polyphenols as dietary supplements, the present study investigated the safety of a standardized polyphenolic extract of clove buds (Clovinol), as assessed by oral acute (5 g/kg b.wt. for 14 days) and subchronic (0.25, 0.5 and...

  1. Managing a massacre: savagery, civility, and gender in Moro Province in the wake of Bud Dajo.

    Science.gov (United States)

    Hawkins, Michael C

    2011-01-01

    This article examines the delicate ideological maneuverings that shaped American colonial constructions of savagery, civility, and gender in the wake of the Bud Dajo massacre in the Philippines's Muslim south in 1906. It looks particularly at shifting notions of femininity and masculinity as these related to episodes of violence and colonial control. The article concludes that, while the Bud Dajo massacre was a terrible black mark on the American military's record in Mindanao and Sulu, colonial officials ultimately used the event to positively affirm existing discourses of power and justification, which helped to sustain and guide military rule in the Muslim south for another seven years.

  2. Stages of flower bud development in Iris pumila and between-habitat morphological differences

    Directory of Open Access Journals (Sweden)

    Barišić-Klisarić Nataša

    2012-01-01

    Full Text Available Previous studies revealed significant phenotypic plasticity and between-population differentiation in flower morphometric traits of Iris pumila in response to environmental variability between natural shade and exposed habitats. Since these habitats differed in flowering times as well, in this work we investigated at which stages of flower bud development differences between open and shaded habitats start to appear. Our analysis detected several groups of trait development patterns through the I. pumila bud development in two contrasting habitats, with stem length being the most suitable trait for application in further analyses of so-called “shade avoidance syndrome���.

  3. Ustawa budżetowa na 2015 rok oraz propozycje zmian układu ustawy budżetowej w Polsce

    OpenAIRE

    Borodo, Andrzej

    2015-01-01

    The article presents the legal content and system of the 2015 Budget Act and the legal significance of its regulations. In the paper there are also proposals to the new structure Polish Budget Act. The present system of the Budget Act does not include the revenues and expenditure of certain state funds (especially the National Road Fund) and finances of State Treasury companies. The current system weakens the control function and budgetary power of the Parliament. New Polish system of the Bud...

  4. The Role of Leaves in Photocontrol of Flower Bud Abscission in Hibiscus rosa-sinensis L. 'Nairobi'

    NARCIS (Netherlands)

    Meeteren, van U.; Gelder, van A.

    2000-01-01

    When compared with exposure to darkness, exposing Hibiscus rosa-sinensis L. 'Nairobi' plants to red light (635 to 685 nm, 2.9 μmol?m-2?s-1) delayed flower bud abscission, while exposure to far-red light (705 to 755 nm, μmol?m-2?s-1) accelerated this process. Flower bud abscission in response to

  5. A unique approach to demonstrating that apical bud temperature specifically determines leaf initiation rate in the dicot Cucumis sativus

    NARCIS (Netherlands)

    Savvides, Andreas; Dieleman, Anja; Ieperen, van Wim; Marcelis, Leo F.M.

    2016-01-01

    Main conclusion: Leaf initiation rate is largely determined by the apical bud temperature even when apical bud temperature largely deviates from the temperature of other plant organs.We have long known that the rate of leaf initiation (LIR) is highly sensitive to temperature, but previous studies

  6. Use of a combination of CEA and tumor budding to identify high-risk patients with stage II colon cancer.

    Science.gov (United States)

    Du, Changzheng; Xue, Weicheng; Dou, Fangyuan; Peng, Yifan; Yao, Yunfeng; Zhao, Jun; Gu, Jin

    2017-07-24

    High-risk patients with stage II colon cancer may benefit from adjuvant chemotherapy, but identifying this patient population can be difficult. We assessed the prognosis value for predicting tumor progression in patients with stage II colon cancer, of a panel of 2 biomarkers for colon cancer: tumor budding and preoperative carcinoembryonic antigen (CEA). Consecutive patients (N = 134) with stage II colon cancer who underwent curative surgery from 2000 to 2007 were included. Multivariate analysis was used to evaluate the association of CEA and tumor budding grade with 5-year disease-free survival (DFS). The prognostic accuracy of CEA, tumor budding grade and the combination of both (CEA-budding panel) was determined. The study found that both CEA and tumor budding grade were associated with 5-year DFS. The prognostic accuracy for disease progression was higher for the CEA-budding panel (82.1%) than either CEA (70.9%) or tumor budding grade (72.4%) alone. The findings indicate that the combination of CEA levels and tumor budding grade has greater prognostic value for identifying patients with stage II colon cancer who are at high-risk for disease progression, than either marker alone.

  7. Expression of the hyaluronan-mediated motility receptor RHAMM in tumor budding cells identifies aggressive colorectal cancers.

    Science.gov (United States)

    Koelzer, Viktor Hendrik; Huber, Bettina; Mele, Valentina; Iezzi, Giandomenica; Trippel, Mafalda; Karamitopoulou, Eva; Zlobec, Inti; Lugli, Alessandro

    2015-11-01

    Expression of the hyaluronan-mediated motility receptor (RHAMM, CD168) predicts adverse clinicopathological features and decreased survival for colorectal cancer (CRC) patients. Using full tissue sections, we investigated the expression of RHAMM in tumor budding cells of 103 primary CRCs to characterize the biological processes driving single-cell invasion and early metastatic dissemination. RHAMM expression in tumor buds was analyzed with clinicopathological data, molecular features and survival. Tumor budding cells at the invasive front of CRC expressed RHAMM in 68% of cases. Detection of RHAMM-positive tumor budding cells was significantly associated with poor survival outcome (P = .0312), independent of TNM stage and adjuvant therapy in multivariate analysis (P = .0201). RHAMM-positive tumor buds were associated with frequent lymphatic invasion (P = .0007), higher tumor grade (P = .0296), and nodal metastasis (P = .0364). Importantly, the prognostic impact of RHAMM expression in tumor buds was maintained independently of the number of tumor buds found in an individual case (P = .0246). No impact of KRAS/BRAF mutation, mismatch repair deficiency and CpG island methylation was observed. RHAMM expression identifies an aggressive subpopulation of tumor budding cells and is an independent adverse prognostic factor for CRC patients. These data support ongoing efforts to develop RHAMM as a target for precision therapy. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. Decreased miR-320a promotes invasion and metastasis of tumor budding cells in tongue squamous cell carcinoma.

    Science.gov (United States)

    Xie, Nan; Wang, Cheng; Zhuang, Zehang; Hou, Jinson; Liu, Xiqiang; Wu, Yue; Liu, Haichao; Huang, Hongzhang

    2016-10-04

    We aimed to determine the specific miRNA profile of tumor budding cells and investigate the potential role of miR-320a in invasion and metastasis of tongue squamous cell carcinoma (TSCC). We collected tumor budding cells and paired central tumor samples from five TSCC specimens with laser capture microdissection and examined the specimens using a miRNA microarray. The specific miRNA signature of tumor budding cells was identified. We found that miR-320a was dramatically decreased in tumor budding cells. Knockdown of miR-320a significantly enhanced migration and invasion of TSCC cell lines. Suz12 was shown to be a direct target of miR-320a. Similar results were also observed in nude mouse models. Multivariate analysis indicated that miR-320a was an independent prognostic factor. Kaplan-Meier analysis demonstrated that decreased miR-320a and high intensity of tumor budding were correlated with poor survival rate, especially in the subgroup with high-intensity tumor budding and low expression of miR-320a. We concluded that decreased expression of miR-320a could promote invasion and metastasis of tumor budding cells by targeting Suz12 in TSCC. A combination of tumor budding and miR-320a may serve as an index to identify an aggressive sub-population of TSCC cells with high metastatic potential.

  9. Correlation of epiphyllous bud differentiation with foliar senescence in crassulacean succulent Kalanchoe pinnata as revealed by thidiazuron and ethrel application.

    Science.gov (United States)

    Jaiswal, Sarita; Sawhney, Sudhir

    2006-05-01

    Leaves of Kalanchoe pinnata have crenate margins with each notch bearing a dormant bud competent to develop into a healthy plantlet. Leaf detachment is a common signal for inducing two contrastingly different leaf-based processes, i.e. epiphyllous bud development into plantlet and foliar senescence. To investigate differentiation of bud and its correlation, if any, with foliar senescence, thidiazuron (TDZ), having cytokinin activity and ethrel (ETH), an ethylene releasing compound, were employed. The experimental system was comprised of marginal leaf discs, each harbouring an epiphyllous bud. Most of the growth characteristics of plantlet developing from the epiphyllous bud were significantly inhibited by TDZ but promoted by ETH. The two regulators modulated senescence in a manner different for leaf discs and plantlet leaves. Thus, TDZ caused a complete retention whereas ETH a complete loss of chlorophyll in the leaf discs. In contrast, the former resulted in a complete depletion of chlorophyll from the plantlet leaves producing an albino effect, while the latter reduced it by 50% only. In combined dispensation of the two regulators, the effect of TDZ was expressed in majority of responses studied. The results presented in this investigation clearly show that the foliar processes of epiphyllous bud differentiation and senescence are interlinked as TDZ that delayed senescence inhibited epiphyllous bud differentiation and ETH that hastened senescence promoted it. A working hypothesis to interpret responsiveness of the disc-bud composite on lines of a source-sink duo, has been proposed.

  10. Dehydration and osmotic adjustment in apple stem tissue during winter as it relates to the frost resistance of buds.

    Science.gov (United States)

    Pramsohler, Manuel; Neuner, Gilbert

    2013-08-01

    In deciduous trees, measurement of stem water potential can be difficult during the leafless period in winter. By using thermocouple psychrometry, osmotic water potentials (Ψo; actual Ψo: Ψo(act); Ψo at full saturation: Ψo(sat)) of expressed sap of bark and bud tissue were measured in order to test if the severity of winter desiccation in apple stems could be sufficiently assessed with Ψo. Water potentials were related to frost resistance and freezing behaviour of buds. The determination of Ψo reliably allowed winter desiccation and osmotic adjustments in apple stem tissue to be assessed. In winter in bark tissue, a pronounced decrease in Ψo(act) and Ψo(sat) was found. Decreased Ψo(sat) indicates active osmotic adjustment in the bark as observed earlier in the leaves of evergreen woody plants. In terminal bud meristems, no significant osmotic adjustments occurred and dehydration during winter was much less. Osmotic water potentials, Ψo(act) and Ψo(sat), of bud tissue were always less negative than in the bark. To prevent water movement and dehydration of the bud tissue via this osmotic gradient, it must be compensated for either by a sufficiently high turgor pressure (Ψp) in bark tissue or by the isolation of the bud tissue from the bark during midwinter. During freezing of apple buds, freeze dehydration and extra-organ freezing could be demonstrated by significantly reduced Ψo(act) values of bud meristems that had been excised in the frozen state. Infrared video thermography was used to monitor freezing patterns in apple twigs. During extracellular freezing of intact and longitudinally dissected stems, infrared differential thermal analysis (IDTA) images showed that the bud meristem remains ice free. Even if cooled to temperatures below the frost-killing temperature, no freezing event could be detected in bud meristems during winter. In contrast, after bud break, terminal buds showed a second freezing at the frost-killing temperature that indicates

  11. Unequivocal Identification of 1-Phenylethyl Acetate in Clove Buds (syzygium aromaticum (L.) Merr. & L.M.Perry) and Clove Essential Oil.

    Science.gov (United States)

    Gassenmeier, Klaus; Schwager, Hugo; Houben, Eric; Clery, Robin

    2017-06-27

    The natural occurrence of 1-phenylethyl acetate (styrallyl acetate) was confirmed in commercially available dried clove buds and also in the hydrodistilled oil from clove buds. This confirms previous reports and other anecdotal evidence for its occurrence in nature.

  12. Unequivocal Identification of 1-Phenylethyl Acetate in Clove Buds (syzygium aromaticum (L.) Merr. & L.M.Perry) and Clove Essential Oil

    OpenAIRE

    Gassenmeier, Klaus; Schwager, Hugo; Houben, Eric; Clery, Robin

    2017-01-01

    The natural occurrence of 1-phenylethyl acetate (styrallyl acetate) was confirmed in commercially available dried clove buds and also in the hydrodistilled oil from clove buds. This confirms previous reports and other anecdotal evidence for its occurrence in nature.

  13. Effect of thawing time, cooling rate and boron nutrition on freezing point of the primordial shoot in norway spruce buds.

    Science.gov (United States)

    Räisänen, Mikko; Repo, Tapani; Lehto, Tarja

    2006-04-01

    Effects of cooling rates on bud frost hardiness have been studied but there is little information on bud responses to thawing. Since the cell wall pore size has been found to increase with boron (B) deficiency, B deficiency may affect the supercooling ability of buds in winter. The effects of duration of thawing time and rate of cooling on bud frost hardiness of Norway spruce (Picea abies) were studied in a B fertilization trial in February 2003 and March 2005. Frost hardiness of apical buds was determined by differential thermal analysis (DTA) and visual scoring of damage. In 2003, the freezing point of primordial shoots of buds (T(f)), i.e. the low-temperature exotherm (LTE), was, on average, -39 degrees C when buds were thawed for less than 3 h and the T(f) increased to -21 degrees C after 18 h of thawing. During the first 4 h of thawing, the rate of dehardening was 6 degrees C h(-1). In 2005, buds dehardened linearly from -39 degrees C to -35 degrees C at a rate of 0.7 degrees C h(-1). In 2003, different cooling rates of 1-5 degrees C h(-1) had a minor effect on T(f) but in 2005 with slow cooling rates T(f) decreased. In both samplings, at cooling rates of 2 and 1 degrees C h(-1), T(f) was slightly higher in B-fertilized than in non-fertilized trees. By contrast, at very short thawing times in 2003, T(f) was somewhat lower in B-fertilized trees. There was little evidence of reduced frost hardiness in trees with low B status. This study showed that buds deharden rapidly when exposed to above-freezing temperatures in winter, but if cooled again they reharden more slowly. According to this study, rapid dehardening of buds has to be taken into account in assessments of frost hardiness.

  14. Tumor budding is associated with an increased risk of lymph node metastasis and poor prognosis in superficial esophageal adenocarcinoma.

    Science.gov (United States)

    Landau, Michael S; Hastings, Steven M; Foxwell, Tyler J; Luketich, James D; Nason, Katie S; Davison, Jon M

    2014-12-01

    The treatment approach for superficial (stage T1) esophageal adenocarcinoma critically depends on the pre-operative assessment of metastatic risk. Part of that assessment involves evaluation of the primary tumor for pathologic characteristics known to predict nodal metastasis: depth of invasion (intramucosal vs submucosal), angiolymphatic invasion, tumor grade, and tumor size. Tumor budding is a histologic pattern that is associated with poor prognosis in early-stage colorectal adenocarcinoma and a predictor of nodal metastasis in T1 colorectal adenocarcinoma. In a retrospective study, we used a semi-quantitative histologic scoring system to categorize 210 surgically resected, superficial (stage T1) esophageal adenocarcinomas according to the extent of tumor budding (none, focal, and extensive) and also evaluated other known risk factors for nodal metastasis, including depth of invasion, angiolymphatic invasion, tumor grade, and tumor size. We assessed the risk of nodal metastasis associated with tumor budding in univariate analyses and controlled for other risk factors in a multivariate logistic regression model. In all, 41% (24 out of 59) of tumors with extensive tumor budding (tumor budding in ≥3 20X microscopic fields) were metastatic to regional lymph nodes, compared with 10% (12 out of 117) of tumors with no tumor budding, and 15% (5 out of 34) of tumors with focal tumor budding (Ptumor budding remains an independent risk factor for lymph node metastasis in superficial esophageal adenocarcinoma associated with a 2.5-fold increase (95% CI=1.1-6.3, P=0.039) in the risk of nodal metastasis. Extensive tumor budding is also a poor prognostic factor with respect to overall survival and time to recurrence in univariate and multivariate analyses. As an independent risk factor for nodal metastasis and poor prognosis after esophagectomy, tumor budding should be evaluated in superficial (T1) esophageal adenocarcinoma as a part of a comprehensive pathologic risk

  15. Characterization of the immunophenotype of the tumor budding and its prognostic implications in squamous cell carcinoma of the lung.

    Science.gov (United States)

    Taira, Tetsuhiko; Ishii, Genichiro; Nagai, Kanji; Yoh, Kiyotaka; Takahashi, Yusuke; Matsumura, Yuki; Kojima, Motohiro; Ohmatsu, Hironobu; Goto, Koichi; Niho, Seiji; Takashima, Hiroshi; Inoue, Hiromasa; Ohe, Yuichiro; Ochiai, Atsushi

    2012-06-01

    Tumor budding is morphologically defined as infiltration by small clusters of cancer cells. While the biological properties of budding cells in adenocarcinoma (decreased expression of adhesion molecules and of differentiation markers) have been elucidated, those of the cells in squamous cell carcinoma (SqCC) of the lung still remain to be clarified. We examined the clinicopathological data of 217 patients with SqCC of the lung. Furthermore we evaluated the immunohistochemical properties of the budding cells. Tumor budding was observed in 83 (38.2%) patients. A statistically significant difference was observed in overall 5-year survival rates between the cases showing tumor budding and the cases not showing budding (45.6% vs. 64.0%, pbudding cells (BCs) exhibited reduced expression levels of the cellular adhesion molecules (E-cadherin; p=0.004, β-catenin; p=0.002) and increased expression levels of laminin-5γ2 (p=0.001). On the other hand, no significant differences in the staining scores for differentiation markers (p63 and podoplanin) were found between BCs and cancer cells forming nests. Multivariate analysis revealed that tumor budding was a significant independent prognostic factor in patients with SqCC of the lung (p=0.022). Tumor budding is an independent adverse prognostic factor in patients with SqCC of the lung. Although budding cells in SqCC exhibited reduced expression levels of the cellular adhesion molecules, the expression levels of specific differentiation markers were retained, suggesting that the budding mechanism in SqCC may differ, at least in part, from that in adenocarcinoma. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  16. A novel minimal in vitro system for analyzing HIV-1 Gag mediated budding

    CERN Document Server

    Gui, Dong; Xu, Jun; Zandi, Roya; Gill, Sarjeet; Huang, I-Chueh; Rao, A L N; Mohideen, Umar

    2013-01-01

    A biomimetic minimalist model membrane is used to study the mechanism and kinetics of the in vitro HIV-1 Gag budding from a giant unilamellar vesicle (GUV). The real time interaction of the Gag, RNA and lipid leading to the formation of minivesicles is measured in real time using confocal microscopy. The Gag is found to lead to resolution limited punctae on the lipid membranes of the GUV. The introduction of the Gag to a GUV solution containing RNA led to the budding of minivesicles on the inside surface of the GUV. The diameter of the GUV decreased due to the bud formation. The corresponding rate of decrease of the GUV diameter was found to be linear in time. The bud formation and the decrease in GUV size were found to be proportional to the Gag concentration. The method is promising and will allow the systematic study of the dynamics of assembly of immature HIV and help classify the hierarchy of factors that impact the Gag protein initiated assembly of retroviruses such as HIV. The GUV system might also be ...

  17. In vitro Proliferation Ability of Axillary Buds in Musa spp | Youmbi ...

    African Journals Online (AJOL)

    As axillary buds have shown mass propagation abilities in Big Ebanga, this explant is tested in other banana varieties to confirm performances already observed. ... Concerning the frequency of leaf emission, the diameter and the height of the pseudostems, no significant differences were obtained between plants derived ...

  18. Mechanisms of taste bud cell loss after head and neck irradiation

    Science.gov (United States)

    Nguyen, Ha M.; Reyland, Mary E.; Barlow, Linda A.

    2012-01-01

    Taste loss in human patients following radiotherapy for head and neck cancer is a common and significant problem, but the cellular mechanisms underlying this loss are not understood. Taste stimuli are transduced by receptor cells within taste buds, and like epidermal cells, taste cells are regularly replaced throughout adult life. This renewal relies on a progenitor cells adjacent to taste buds, which continually supply new cells to each bud. Here we treated adult mice with a single 8 Gy dose of X-ray irradiation to the head and neck, and analyzed taste epithelium at 1–21 days post-irradiation (dpi). We found irradiation targets the taste progenitor cells, which undergo cell cycle arrest (1–3 dpi) and apoptosis (within 1 dpi). Taste progenitors resume proliferation at 5–7 dpi, with the proportion of cells in S and M phase exceeding control levels at 5–6 and 6 dpi, respectively, suggesting that proliferation is accelerated and/or synchronized following radiation damage. Using BrdU birthdating to identify newborn cells, we found that the decreased proliferation following irradiation reduces the influx of cells at 1–2 dpi, while the robust proliferation detected at 6 dpi accelerates entry of new cells into taste buds. By contrast, the number of differentiated taste cells was not significantly reduced until 7 dpi. These data suggest a model where continued natural taste cell death, paired with temporary interruption of cell replacement underlies taste loss after irradiation. PMID:22399770

  19. Genes in the Ureteric Budding Pathway : Association Study on Vesico-Ureteral Reflux Patients

    NARCIS (Netherlands)

    van Eerde, Albertien M.; Duran, Karen; van Riel, Els; de Kovel, Carolien G. F.; Koeleman, Bobby P. C.; Knoers, Nine V. A. M.; Renkema, Kirsten Y.; van der Horst, Henricus J. R.; Bokenkamp, Arend; van Hagen, Johanna M.; van den Berg, Leonard H.; Wolffenbuttel, Katja P.; van den Hoek, Joop; Feitz, Wouter F.; de Jong, Tom P. V. M.; Giltay, Jacques C.; Wijmenga, Cisca

    2012-01-01

    Vesico-ureteral reflux (VUR) is the retrograde passage of urine from the bladder to the urinary tract and causes 8.5% of end-stage renal disease in children. It is a complex genetic developmental disorder, in which ectopic embryonal ureteric budding is implicated in the pathogenesis. VUR is part of

  20. Genes in the ureteric budding pathway: Association study on vesico-ureteral reflux patients

    NARCIS (Netherlands)

    A.M. van Eerde (Albertien ); K. Duran (Karen); E. van Riel (Els); C.G.F. de Kovel (Carolien); B.P.C. Koeleman (Bobby); N.V.A.M. Knoers (Nine); K.Y. Renkema (Kirsten); H.J.R. van der Horst (Henricus); A. Bökenkamp (Arend); J.M. van Hagen (Johanna); L.H. van den Berg (Leonard); K.P. Wolffenbuttel (Katja); J. van den Hoek (Joop); W.F.J. Feitz (Wout); T.P.V.M. de Jong (Tom); J.C. Giltay (Jacques); C. Wijmenga (Cisca)

    2012-01-01

    textabstractVesico-ureteral reflux (VUR) is the retrograde passage of urine from the bladder to the urinary tract and causes 8.5% of end-stage renal disease in children. It is a complex genetic developmental disorder, in which ectopic embryonal ureteric budding is implicated in the pathogenesis. VUR

  1. Genes in the ureteric budding pathway: association study on vesico-ureteral reflux patients.

    NARCIS (Netherlands)

    Eerde, A.M. van; Duran, K.; Riel, E. van; Kovel, C.G.F. de; Koeleman, B.P.; Knoers, N.V.A.M.; Renkema, K.Y.; Horst, H.J. van der; Bokenkamp, A.; Hagen, J.M. van; Berg, L.H. van den; Wolffenbuttel, K.P.; Hoek, J. van den; Feitz, W.F.J.; Jong, T.P. de; Giltay, J.C.; Wijmenga, C.

    2012-01-01

    Vesico-ureteral reflux (VUR) is the retrograde passage of urine from the bladder to the urinary tract and causes 8.5% of end-stage renal disease in children. It is a complex genetic developmental disorder, in which ectopic embryonal ureteric budding is implicated in the pathogenesis. VUR is part of

  2. Genes in the Ureteric Budding Pathway: Association Study on Vesico-Ureteral Reflux Patients

    NARCIS (Netherlands)

    van Eerde, Albertien M.; Duran, Karen; van Riel, Els; de Kovel, Carolien G. F.; Koeleman, Bobby P. C.; Knoers, Nine V. A. M.; Renkema, Kirsten Y.; van der Horst, Henricus J. R.; Bökenkamp, Arend; van Hagen, Johanna M.; van den Berg, Leonard H.; Wolffenbuttel, Katja P.; van den Hoek, Joop; Feitz, Wouter F.; de Jong, Tom P. V. M.; Giltay, Jacques C.; Wijmenga, Cisca

    2012-01-01

    Vesico-ureteral reflux (VUR) is the retrograde passage of urine from the bladder to the urinary tract and causes 8.5% of end-stage renal disease in children. It is a complex genetic developmental disorder, in which ectopic embryonal ureteric budding is implicated in the pathogenesis. VUR is part of

  3. Biology and behavior of a larch bud moth, Zeiraphera sp., in Alaska.

    Science.gov (United States)

    Richard A. Werner

    1980-01-01

    A possibly new species or subspecies of larch bud moth of the genus Zeiraphera, closely related to Z. improbana (Walker), was found associated with tamarack, Larix laricina (Du Roi) K. Koch, stands in interior Alaska. An outbreak occurred during 1975 and 1976 over an area of 240 000 ha (590,000 acres)....

  4. Hormonal influence on the in vitro bud burst of some cassava ...

    African Journals Online (AJOL)

    This work tested the effect of different growth regulators on Manihot esculenta explants cultured. Nodal segments were disinfected and cultivated on Murashige and Skoog's basal media. The effects of the different combinations on bud burst were observed after five weeks of culture. The results show an interaction between ...

  5. Effect of pre-budwood collection treatment on budding success in ...

    African Journals Online (AJOL)

    The inconsistencies in success of budding exercises pose great problem in the rapid multiplication of desirable genotypes for germplasm, clonal plot and seed garden establishment in cacao, Theobroma cacao L. This study was carried out at Cocoa Research Institute of Nigeria, Ibadan to determine the effects of ...

  6. Effect of floral bud reduction on flower longevity in Asiatic hybrids lilies.

    NARCIS (Netherlands)

    Meulen-Muisers, van der J.J.M.; Oeveren, van J.C.; Sandbrink, J.M.; Tuyl, van J.M.

    1995-01-01

    Floral bud abortion was found to be an undesirable source of non-genetic variation in breeding trials directed on the improvement of individual flower longevity in Asiatic hybrid lilies. It increased the longevity of the remaining flowers of the inflorescence. A similar response was found after

  7. Hormonal influence on the in vitro bud burst of some cassava ...

    African Journals Online (AJOL)

    KGILLES

    2013-03-27

    Mar 27, 2013 ... Full Length Research Paper. Hormonal influence on the in vitro bud burst of some ... export culture after cotton, coffee, palm tree which pro- vides substantial incomes to producers. Cassava holds a .... The explants were washed under tap water for 10 min. Ten node cuttings from young stems of each of the ...

  8. Reproductive effects of lipid soluble components of Syzygium aromaticum flower bud in male mice

    Directory of Open Access Journals (Sweden)

    Raghav Kumar Mishra

    2013-01-01

    Full Text Available Background: The flower buds of Syzygium aromaticum (clove have been used in indigenous medicines for the treatment of male sexual disorders in Indian subcontinent. Objective: To evaluate the effect of Syzygium aromaticum flower bud on male reproduction, using Parkes (P strain mice as animal model. Materials and Methods: Mice were orally administered lipid soluble components of Syzygium aromaticum flower bud in doses of 15, 30, and 60 mg/kg body weight for 35 days, and several male reproductive endpoints were evaluated. Results: Treatment with lower dose (15 mg of Syzygium increased the motility of sperm and stimulated the secretory activities of epididymis and seminal vesicle, while higher doses (30 and 60 mg had adverse effects on sperm dynamics of cauda epididymidis and on the secretory activities of epididymis and seminal vesicle. Libido was not affected in treated males; however, a significant decrease in litter in females sired by males treated with higher doses of Syzygium was recorded. Conclusion: Treatment with Syzygium aromaticum flower bud causes dose-dependent biphasic effect on male reproductive indices in P mice; lower dose of Syzygium appears stimulatory, while the higher doses have adverse effect on male reproduction. The results suggest that the lower dose of Syzygium may have androgenic effect, but further studies are needed to support this contention.

  9. Viral and cellular requirements for the budding of feline endogenous retrovirus RD-114.

    Science.gov (United States)

    Fukuma, Aiko; Abe, Masumi; Urata, Shuzo; Yoshikawa, Rokusuke; Morikawa, Yuko; Miyazawa, Takayuki; Yasuda, Jiro

    2011-12-14

    RD-114 virus is a feline endogenous retrovirus and produced as infectious viruses in some feline cell lines. Recently, we reported the contamination of an infectious RD-114 virus in a proportion of live attenuated vaccines for dogs and cats. It is very difficult to completely knock out the RD-114 proviruses from cells, as endogenous retroviruses are usually integrated multiply into the host genome. However, it may be possible to reduce the risk of contamination of RD-114 virus by regulating the viral release from cells. In this study, to understand the molecular mechanism of RD-114 virus budding, we attempted to identify the viral and cellular requirements for RD-114 virus budding. Analyses of RD-114 L-domain mutants showed that the PPPY sequence in the pp15 region of Gag plays a critical role in RD-114 virus release as viral L-domain. Furthermore, we investigated the cellular factors required for RD-114 virus budding. We demonstrated that RD-114 virus release was inhibited by overexpression of dominant negative mutants of Vps4A, Vps4B, and WWP2. These results strongly suggest that RD-114 budding utilizes the cellular multivesicular body sorting pathway similar to many other retroviruses.

  10. In vitro culture techniques as a tool of sugarcane bud germination ...

    African Journals Online (AJOL)

    Germination was the first stage confronted to soil salinity and it is important to determine salt effects on this stage. In this study, we reported an in vitro procedure for studying sugarcane bud germination and shoot growth under salt stress with different NaCl concentrations (0, 17, 34, 68 and 102 mM) using cultivar NCo310.

  11. Safety assessment of Syzygium aromaticum flower bud (clove) extract with respect to testicular function in mice.

    Science.gov (United States)

    Mishra, Raghav Kumar; Singh, Shio Kumar

    2008-10-01

    The flower buds of Syzygium aromaticum (clove), a common food flavor, have been used as indigenous medicine for the treatment of male sexual disorders in Asian countries. However, the possible mechanism(s) by which it acts at testicular level remain obscure. Therefore, to investigate its effect on testicular function, chronic oral exposure of hexane extract of flower buds of Syzygium aromaticum in three doses (15 mg, 30 mg, and 60 mg/kg BW) were evaluated for a single spermatogenic cycle (35 days) in Parkes (P) strain mice. The treatment did not induce systemic toxicity at the doses tested. Lower dose (15 mg) of the extract increased the activities of Delta(5) 3 beta-HSD and 17 beta-HSD, and serum level of testosterone. The higher doses (30 and 60 mg) of extract inhibited these parameters and induced non-uniform degenerative changes in the seminiferous tubules associated with decrease in daily sperm production and depletion of 1C (round and elongated spermatids) population. Taken together these results suggest biphasic action of hexane extract of Syzygium aromaticum flower bud on testicular function, thereby advocating a cautious use of the flower bud as an aphrodisiac in indigenous systems of medicine in Asian countries.

  12. Somatic embryogenesis in cv. Navolean using shoots apexes from axillary buds

    Directory of Open Access Journals (Sweden)

    Jorge López

    2005-04-01

    Full Text Available In order to develop embryogenic cultures in AAB Musa genotypes without persistent male inflorescence, the process has had greater success from proliferating meristems for callus formation with embryogenic structures. Based on the previous information, other alternative explant sources for somatic embryogenesis development in cv. Navolean. Meristematic apexes were cultured in p5 culture medium supplemented with thidiazuron and ancymidol (0.2, 0.4, 0.6, mg.l-1 to obtain axillary buds. Later, axillary buds and proliferated meristems were tested for callus induction with embryogenic structures combinations with different 2,4-D concentrations. The best growth regulator for obtaining axillary buds was ancymidol (0.2 mg.l-1. For callus formation with embryogenic structures, axillary buds at 1.0 mg.l-1 2,4-D provided a higher percentage (13.6%. These results permitted the development of embryogenic cell suspensions from somatic embryos. Key words: Ancymidol, embryogenic cell suspension, plantain

  13. Surface disinfection procedure and in vitro regeneration of grapevine (Vitis vinifera L.) axillary buds.

    Science.gov (United States)

    Lazo-Javalera, M F; Troncoso-Rojas, R; Tiznado-Hernández, M E; Martínez-Tellez, M A; Vargas-Arispuro, I; Islas-Osuna, M A; Rivera-Domínguez, M

    2016-01-01

    Establishment of an efficient explants surface disinfection protocol is essential for in vitro cell and tissue culture as well as germplasm conservation, such as the case of Grapevine (Vitis spp.) culture. In this research, different procedures for disinfection and regeneration of field-grown grapevine cv. 'Flame seedless' axillary buds were evaluated. The buds were disinfected using either NaOCl or allyl, benzyl, phenyl and 2-phenylethyl isothiocyanates. Two different media for shooting and four media for rooting were tested. Shoot and root development per buds were registered. The best disinfection procedure with 90 % of tissue survival involved shaking for 60 min in a solution containing 20 % Clorox with 50 drops/L Triton(®) X-100. These tissues showed the potential to regenerate a complete plant. Plant regeneration was conducted using full strength Murashigue and Skoog (MS) medium supplemented with 8 µM benzyl aminopurine for shoot induction and multiplication, whereas rooting was obtained on half strength MS supplemented with 2 mg L(-1) of indole-3-butyric acid and 200 mg L(-1) of activated charcoal. In this work, it was designed the protocols for obtaining sterile field-grown grapevine buds and in vitro plant development. This methodology showed potential to produce vigorous and healthy plants in 5 weeks for clonal grapevine propagation. Regenerated plants were successfully established in soil.

  14. Continuous High-resolution Microscopic Observation of Replicative Aging in Budding Yeast

    NARCIS (Netherlands)

    Huberts, Daphne H. E. W.; Janssens, Georges E.; Lee, Sung Sik; Vizcarra, Ima Avalos; Heinemann, Matthias

    We demonstrate the use of a simple microfluidic setup, in which single budding yeast cells can be tracked throughout their entire lifespan. The microfluidic chip exploits the size difference between mother and daughter cells using an array of micropads. Upon loading, cells are trapped underneath

  15. Whole lifespan microscopic observation of budding yeast aging through a microfluidic dissection platform

    NARCIS (Netherlands)

    Lee, Sung Sik; Avalos Vizcarra, Ima; Huberts, Daphne H E W; Lee, Luke P; Heinemann, Matthias

    2012-01-01

    Important insights into aging have been generated with the genetically tractable and short-lived budding yeast. However, it is still impossible today to continuously track cells by high-resolution microscopic imaging (e.g., fluorescent imaging) throughout their entire lifespan. Instead, the field

  16. Gall mite inspection on dormant black currant buds using machine vision

    DEFF Research Database (Denmark)

    Nielsen, M. R.; Stigaard Laursen, Morten; Jonassen, M. S.

    2013-01-01

    This paper presents a novel machine vision-based approach detecting and mapping gall mite infection in dormant buds on black currant bushes. A vehicle was fitted with four cameras and RTK-GPS. Results compared automatic detection to human decisions based on the images, and by mapping the results ...

  17. Deltabaculoviruses encode a functional type I budded virus envelope fusion protein

    Science.gov (United States)

    Envelope fusion proteins (F proteins) are major constituents of budded viruses (BVs) of alpha- and betabaculoviruses (Baculoviridae) and are essential for the systemic infection of insect larvae and insect cells in culture. An F protein homolog gene was absent in gammabaculoviruses. Here we show tha...

  18. Conservation of the USDA-ARS National Plant Germplasm System Apple Collection using dormant bud cryopreservation

    Science.gov (United States)

    The USDA-ARS National Plant Germplasm System maintains a grafted collection of apple accessions representing 49 taxa in Geneva, NY. Since 1993, dormant buds of many of these accessions have been routinely cryopreserved at the USDA-ARS National Center for Genetic Resources Preservation (NCGRP) in For...

  19. Buds of Parenting in Emerging Adult Males: What We Learned from Our Parents

    Science.gov (United States)

    Scharf, Miri; Mayseless, Ofra

    2011-01-01

    The authors examine the precursors of parenting buds (representations regarding parenting before actual parenting) by following 60 men from adolescence to emerging adulthood. Quality of relationships with parents, and attachment representations (state of mind with respect to attachment and attachment styles) assessed in adolescence, contribute to…

  20. Dehydration and vernalization treatments identify overlapping molecular networks impacting endodormancy maintenance in leafy spurge crown buds

    Science.gov (United States)

    Leafy spurge (Euphorbia esula L.) is an herbaceous perennial weed that reproduces vegetatively from an abundance of underground adventitious buds (UABs), which undergo well-defined phases of seasonal dormancy (para-, endo- and eco-dormancy). In this study, the effects of dehydration-stress on vegeta...

  1. In vitro culture techniques as a tool of sugarcane bud germination ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-10-20

    Oct 20, 2008 ... Germination was the first stage confronted to soil salinity and it is important to determine salt effects on this stage. In this study, we reported an in vitro procedure for studying sugarcane bud germination and shoot growth under salt stress with different NaCl concentrations (0, 17, 34, 68 and 102 mM) using.

  2. Dehydration improves cryopreservation of mat rush (Juncus decipiens Nakai) basal stem buds on cryo-plates.

    Science.gov (United States)

    Niino, T; Yamamoto, S I; Fukui, K; Castillo Martinez, C R; Arizaga, M V; Matsumoto, T; Engelmann, F

    2013-01-01

    Two cryopreservation procedures using aluminium cryo-plates, termed V-Cryo-plate and D-Cryo-plate, were successfully developed for in vitro mat rush (Juncus decipiens Nakai) basal stem buds. Multiple stems induced in liquid MS medium containing 8.9 μM BA by roller culture were cut into small clumps, plated on solid MS medium and cultured for 1 week at 25 degree C. Clumps that had produced many buds were cold-hardened at 5 degree C for 1-2 months. The buds with basal stems were dissected from small clumps and precultured overnight at 25 degree C on solid MS medium containing 0.3 M sucrose. Precultured buds were placed on aluminium cryo-plates and embedded in calcium alginate gel. Osmoprotection was performed by immersing the cryo-plates for 30 min at 25 degree C in loading solution (2 M glycerol + 1.0 M sucrose). In the D-Cryo-plate procedure, the buds were dehydrated to 27-25% moisture content (fresh weight) by placing the cryo-plates in the air current of a laminar flow cabinet for 2 to 3 h. In the V-Cryo-plate procedure, buds were dehydrated by immersing the cryo-plates in PVS2 vitrification solution for 40 min at 25 degree C. In both procedures, cooling was performed by placing the cryo-plates in uncapped cryotubes, which were immersed in liquid nitrogen. For rewarming, cryo-plates were immersed in medium with 1.0 M sucrose for 20 min at room temperature. Regrowth of cryopreserved buds of line 'Kitakei 2' using D-Cryo-plate and V-Cryo-plate procedures, was 90% and 80%, respectively. The two procedures were applied to 20 additional mat rush lines. Using the V-Cryo-plate procedure resulted in regrowth ranging between 13.3 and 86.7%, with an average of 52.5%. The D-Cryo-plate led to regrowth ranging between 73.3 and 96.7%, with an average of 86.3%. The D-Cryo-plate procedure will facilitate cryostorage of mat rush germplasm.

  3. Tumor budding in colorectal cancer revisited: results of a multicenter interobserver study.

    Science.gov (United States)

    Koelzer, Viktor H; Zlobec, Inti; Berger, Martin D; Cathomas, Gieri; Dawson, Heather; Dirschmid, Klaus; Hädrich, Marion; Inderbitzin, Daniel; Offner, Felix; Puppa, Giacomo; Seelentag, Walter; Schnüriger, Beat; Tornillo, Luigi; Lugli, Alessandro

    2015-05-01

    Tumor budding in colorectal cancer (CRC) is recognized as a valuable prognostic factor but its translation into daily histopathology practice has been delayed by lack of agreement on the optimal method of assessment. Within the context of the Swiss Association of Gastrointestinal Pathology (SAGIP), we performed a multicenter interobserver study on tumor budding, comparing hematoxylin and eosin (H&E) with pan-cytokeratin staining using a 10 high power field (10HPF) and hotspot (1HPF) method. Two serial sections of 50 TNM stage II-IV surgically treated CRC were stained for H&E and pan-cytokeratin. Tumor buds were scored by independent observers at six participating centers in Switzerland and Austria using the 10HPF and 1HPF method on a digital pathology platform. Pearson correlation (r) and intra-class correlation coefficients (ICC) comparing scores between centers were calculated. Three to four times more tumor buds were detected in pan-cytokeratin compared to H&E slides. Correlation coefficients for tumor budding counts between centers ranged from r = 0.46 to r = 0.91 for H&E and from r = 0.73 to r = 0.95 for pan-cytokeratin slides. Interobserver agreement across all centers was excellent for pan-cytokeratin [10HPF: ICC = 0.83 and 1HPF: ICC = 0.8]. In contrast, assessment of tumor budding on H&E slides reached only moderate agreement [10HPF: ICC = 0.58 and 1HPF: ICC = 0.49]. Based on previous literature and our findings, we recommend (1) pan-cytokeratin staining whenever possible, (2) 10HPF method for resection specimens, and (3) 1HPF method for limited material (preoperative biopsy or pT1). Since tumor budding counts can be used to determine probabilities of relevant outcomes and as such more optimally complement clinical decision making, we advocate the avoidance of cutoff scores.

  4. Prognostic significance of tumor budding and single cell invasion in gastric adenocarcinoma

    Directory of Open Access Journals (Sweden)

    Che K

    2017-02-01

    Full Text Available Keying Che,1,* Yang Zhao,2,3,* Xiao Qu,1 Zhaofei Pang,1 Yang Ni,4 Tiehong Zhang,4 Jiajun Du,1,5 Hongchang Shen4 1Institute of Oncology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, 2Department of Breast Surgery, Key Laboratory of Breast Cancer in Shanghai, Collaborative Innovation Center of Cancer Medicine, Fudan University Shanghai Cancer Center, 3Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, 4Department of Oncology, Shandong Provincial Hospital Affiliated to Shandong University, 5Department of Thoracic Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, People’s Republic of China *These authors contributed equally to this work Purpose: Gastric carcinoma (GC is a highly aggressive cancer and one of the leading causes of cancer-related deaths worldwide. Histopathological evaluation pertaining to invasiveness is likely to provide additional information in relation to patient outcome. In this study, we aimed to evaluate the prognostic significance of tumor budding and single cell invasion in gastric adenocarcinoma.Materials and methods: Hematoxylin and eosin-stained slides generated from 296 gastric adenocarcinoma patients with full clinical and pathological and follow-up information were systematically reviewed. The patients were grouped on the basis of tumor budding, single cell invasion, large cell invasion, mitotic count, and fibrosis. The association between histopathological parameters, different classification systems, and overall survival (OS was statistically analyzed.Results: Among the 296 cases that were analyzed, high-grade tumor budding was observed in 49.0% (145 of them. Single cell invasion and large cell invasion were observed in 62.8% (186 and 16.9% (50 of the cases, respectively. Following univariate analysis, patients with high-grade tumor budding had shorter OS than those with low-grade tumor budding (hazard ratio [HR]: 2.260, P<0

  5. Mapping of Candidate Genes Involved in Bud Dormancy and Flowering Time in Sweet Cherry (Prunus avium.

    Directory of Open Access Journals (Sweden)

    Sophie Castède

    Full Text Available The timing of flowering in perennial plants is crucial for their survival in temperate climates and is regulated by the duration of bud dormancy. Bud dormancy release and bud break depend on the perception of cumulative chilling during endodormancy and heat during the bud development. The objectives of this work were to identify candidate genes involved in dormancy and flowering processes in sweet cherry, their mapping in two mapping progenies 'Regina' × 'Garnet' and 'Regina' × 'Lapins', and to select those candidate genes which co-localized with quantitative trait loci (QTLs associated with temperature requirements for bud dormancy release and flowering. Based on available data on flowering processes in various species, a list of 79 candidate genes was established. The peach and sweet cherry orthologs were identified and primers were designed to amplify sweet cherry candidate gene fragments. Based on the amplified sequences of the three parents of the mapping progenies, SNPs segregations in the progenies were identified. Thirty five candidate genes were genetically mapped in at least one of the two progenies and all were in silico mapped. Co-localization between candidate genes and QTLs associated with temperature requirements and flowering date were identified for the first time in sweet cherry. The allelic composition of the candidate genes located in the major QTL for heat requirements and flowering date located on linkage group 4 have a significant effect on these two traits indicating their potential use for breeding programs in sweet cherry to select new varieties adapted to putative future climatic conditions.

  6. Knocking out P2X receptors reduces transmitter secretion in taste buds

    Science.gov (United States)

    Huang, Yijen A.; Stone, Leslie M.; Pereira, Elizabeth; Yang, Ruibiao; Kinnamon, John C.; Dvoryanchikov, Gennady; Chaudhari, Nirupa; Finger, Thomas E.; Kinnamon, Sue C.; Roper, Stephen D.

    2011-01-01

    In response to gustatory stimulation, taste bud cells release a transmitter, ATP, that activates P2X2 and P2X3 receptors on gustatory afferent fibers. Taste behavior and gustatory neural responses are largely abolished in mice lacking P2X2 and P2X3 receptors (P2X2 and P2X3 double knockout, or “DKO” mice). The assumption has been that eliminating P2X2 and P2X3 receptors only removes postsynaptic targets but that transmitter secretion in mice is normal. Using functional imaging, ATP biosensor cells, and a cell-free assay for ATP, we tested this assumption. Surprisingly, although gustatory stimulation mobilizes Ca2+ in taste Receptor (Type II) cells from DKO mice, as from wild type (WT) mice, taste cells from DKO mice fail to release ATP when stimulated with tastants. ATP release could be elicited by depolarizing DKO Receptor cells with KCl, suggesting that ATP-release machinery remains functional in DKO taste buds. To explore the difference in ATP release across genotypes, we employed reverse transcriptase (RT)-PCR, immunostaining, and histochemistry for key proteins underlying ATP secretion and degradation: Pannexin1, TRPM5, and NTPDase2 (ecto-ATPase) are indistinguishable between WT and DKO mice. The ultrastructure of contacts between taste cells and nerve fibers is also normal in the DKO mice. Finally, quantitative RT-PCR show that P2X4 and P2X7, potential modulators of ATP secretion, are similarly expressed in taste buds in WT and DKO taste buds. Importantly, we find that P2X2 is expressed in WT taste buds and appears to function as an autocrine, positive feedback signal to amplify taste-evoked ATP secretion. PMID:21940456

  7. Sea buckthorn bud extract displays activity against cell-cultured Influenza virus.

    Science.gov (United States)

    Torelli, A; Gianchecchi, E; Piccirella, S; Manenti, A; Piccini, G; Llorente Pastor, E; Canovi, B; Montomoli, E

    2015-08-05

    Vaccines and antiviral drugs are the most widely used methods of preventing or treating Influenza virus infection. The role of sea buckthorn (SBT) bud dry extract as a natural antiviral drug against Influenza was investigated. Influenza virus was cultured in the MDCK cell line, with or without SBT bud extract, and virus growth was assessed by HA and TCID50 virus titration in terms of cytopathic effect on cells. Several concentrations of extract were tested, the virus titer being measured on day 4 after infection. After infection, the virus titer in the control sample was calculated to be 2.5 TCID50/ml; treatment with SBT bud extract reduced the virus titer to 2.0 TCID50/ml at 50 μg/ml, while the HA titer was reduced from 1431 (control) to 178. Concentrations lower than 50μg/ml displayed an inhibitory effect in the HA assay, but not in the TCID50 virus titration; however, observation of the viral cultures confirmed a slowdown of viral growth at all concentrations. Natural dietary supplements and phytotherapy are a growing market and offer new opportunities for the treatment of several diseases and disorders. These preliminary experiments are the first to show that SBT bud extract is able to reduce the growth of the Influenza A H1N1 virus in vitro at a concentration of 50 μg/ml. This discovery opens up the possibility of using SBT bud extract as a valid weapon against Influenza and, in addition, as the starting-point for the discovery of new drugs. © Copyright by Pacini Editore SpA, Pisa, Italy.

  8. Galectin-3 promotes HIV-1 budding via association with Alix and Gag p6.

    Science.gov (United States)

    Wang, Sheng-Fan; Tsao, Ching-Han; Lin, Yu-Ting; Hsu, Daniel K; Chiang, Meng-Lin; Lo, Chia-Hui; Chien, Fan-Ching; Chen, Peilin; Arthur Chen, Yi-Ming; Chen, Huan-Yuan; Liu, Fu-Tong

    2014-11-01

    Galectin-3 has been reported to regulate the functions of a number of immune cell types. We previously reported that galectin-3 is translocated to immunological synapses in T cells upon T-cell receptor engagement, where it associates with ALG-2-interacting protein X (Alix). Alix is known to coordinate with the endosomal sorting complex required for transport (ESCRT) to promote human immunodeficiency virus (HIV)-1 virion release. We hypothesized that galectin-3 plays a role in HIV-1 viral budding. Cotransfection of cells of the Jurkat T line with galectin-3 and HIV-1 plasmids resulted in increased HIV-1 budding, and suppression of galectin-3 expression by RNAi in Hut78 and primary CD4+ T cells led to reduced HIV-1 budding. We used immunofluorescence microscopy to observe the partial colocalization of galectin-3, Alix and Gag in HIV-1-infected cells. Results from co-immunoprecipitation experiments indicate that galectin-3 expression promotes Alix-Gag p6 association, whereas the results of Alix knockdown suggest that galectin-3 promotes HIV-1 budding through Alix. HIV-1 particles released from galectin-3-expressing cells acquire the galectin-3 protein in an Alix-dependent manner, with proteins primarily residing inside the virions. We also found that the galectin-3 N-terminal domain interacts with the proline-rich region of Alix. Collectively, these results suggest that endogenous galectin-3 facilitates HIV-1 budding by promoting the Alix-Gag p6 association. © The Author 2014. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  9. Vismodegib, an antagonist of hedgehog signaling, directly alters taste molecular signaling in taste buds.

    Science.gov (United States)

    Yang, Hyekyung; Cong, Wei-Na; Yoon, Jeong Seon; Egan, Josephine M

    2015-02-01

    Vismodegib, a highly selective inhibitor of hedgehog (Hh) pathway, is an approved treatment for basal-cell carcinoma. Patients on treatment with vismodegib often report profound alterations in taste sensation. The cellular mechanisms underlying the alterations have not been studied. Sonic Hh (Shh) signaling is required for cell growth and differentiation. In taste buds, Shh is exclusively expressed in type IV taste cells, which are undifferentiated basal cells and the precursors of the three types of taste sensing cells. Thus, we investigated if vismodegib has an inhibitory effect on taste cell turnover because of its known effects on Hh signaling. We gavaged C57BL/6J male mice daily with either vehicle or 30 mg/kg vismodegib for 15 weeks. The gustatory behavior and immunohistochemical profile of taste cells were examined. Vismodegib-treated mice showed decreased growth rate and behavioral responsivity to sweet and bitter stimuli, compared to vehicle-treated mice. We found that vismodegib-treated mice had significant reductions in taste bud size and numbers of taste cells per taste bud. Additionally, vismodegib treatment resulted in decreased numbers of Ki67- and Shh-expressing cells in taste buds. The numbers of phospholipase Cβ2- and α-gustducin-expressing cells, which contain biochemical machinery for sweet and bitter sensing, were reduced in vismodegib-treated mice. Furthermore, vismodegib treatment resulted in reduction in numbers of T1R3, glucagon-like peptide-1, and glucagon-expressing cells, which are known to modulate sweet taste sensitivity. These results suggest that inhibition of Shh signaling by vismodegib treatment directly results in alteration of taste due to local effects in taste buds. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.

  10. Investigation the Frost Resistance of Vegetative and Reproductive Buds of Pear Cultivars in Mashhad Climate Condition

    Directory of Open Access Journals (Sweden)

    shadan khorshidi

    2017-02-01

    Full Text Available Introduction: Most deciduous trees need low temperature to break flower bud dormancy. One of the most important abiotic stresses is low temperature which limits production of temperate fruits. Pear production has been considerably reduced in recent years. Important pear cultivars show different levels of resistance to cold. Cold compatibility followed by resistance increase is controlled genetically and contains several mechanisms which lead to production of different metabolites such as: polypeptides, amino acids and sugars. The object of this research was to evaluate the frost resistance of different ‘Dare Gazi’ genotypes and other pear cultivars in Mashhad climate condition. Materials and Methods: This study was conducted to investigate the frost resistance of 23 ‘Dare Gazi’ pear genotypes and nine other cultivars include: ‘William’s’, ‘Bell de june’, ‘Spadona’, ‘Koshia’, ‘Domkaj’, ‘Torsh’, ‘Sebri’ and ‘Tabrizi’. Plant material contained vegetative and reproductive buds of one-year-old shoot samples which were collected from 25-year old trees on March 2014, four days after winter cold (-6.6 °C in three directions of trees and sent to the laboratory. Frost damages of vegetative and reproductive buds were investigated based on visual observations (%, electrolyte leakage (EC and proline content. EC was measured with a Metrohm 644 digital conductivity meter and proline content was measured based on Bates et al. (1973 method, using acid ninhydrin. The experiment was performed on completely randomized experimental design with three replications. Statistical analysis was carried out using MSTAT-C and Excel software. Mean values were compared using the least significance difference test (LSD at 1% levels. Cluster analysis was conducted by SPSS 16 program. Results and Discussion: Highest EC of reproductive buds was observed in ‘Dare Gazi’ 10, 19, ‘Tabrizi’ and ‘Torsh’ whereas ‘Dare Gazi’ 8, 18

  11. The timing of bud break in warming conditions: variation among seven sympatric conifer species from Eastern Canada

    Science.gov (United States)

    Rossi, Sergio; Isabel, Nathalie

    2017-11-01

    Phenological changes are expected with the ongoing global warming, which could create mismatches in the growth patterns among sympatric species or create synchrony with insect herbivores. In this study, we performed a comparative assessment of the timings of bud break among seven conifer species of Eastern Canada by evaluating seedling development in growth chambers under different temperatures (16, 20 and 24 °C). Bud break occurred earliest in Larix laricina, while Pinus strobus and Pinus resinosa had the latest. Warmer conditions advanced bud break, with the greatest effects being observed at the lower temperatures. Mixed models estimated that one additional degree of temperature produced advancements of 5.3 and 2.1 days at 16 and 20 °C, respectively. The hypothesis of an asynchronous change between species under warming was demonstrated only for the last phenological phases (split buds and exposed shoots), and principally in pines. Abies balsamea showed changes in bud break comparable with the other species analysed, rejecting the hypothesis of mismatches under warmer conditions. The observed non-linear responses of the timings of bud break to warming suggest that the major changes in bud phenology should be expected at the lowest temperatures.

  12. The timing of bud break in warming conditions: variation among seven sympatric conifer species from Eastern Canada

    Science.gov (United States)

    Rossi, Sergio; Isabel, Nathalie

    2017-06-01

    Phenological changes are expected with the ongoing global warming, which could create mismatches in the growth patterns among sympatric species or create synchrony with insect herbivores. In this study, we performed a comparative assessment of the timings of bud break among seven conifer species of Eastern Canada by evaluating seedling development in growth chambers under different temperatures (16, 20 and 24 °C). Bud break occurred earliest in Larix laricina, while Pinus strobus and Pinus resinosa had the latest. Warmer conditions advanced bud break, with the greatest effects being observed at the lower temperatures. Mixed models estimated that one additional degree of temperature produced advancements of 5.3 and 2.1 days at 16 and 20 °C, respectively. The hypothesis of an asynchronous change between species under warming was demonstrated only for the last phenological phases (split buds and exposed shoots), and principally in pines. Abies balsamea showed changes in bud break comparable with the other species analysed, rejecting the hypothesis of mismatches under warmer conditions. The observed non-linear responses of the timings of bud break to warming suggest that the major changes in bud phenology should be expected at the lowest temperatures.

  13. In silico analysis of 3 expansin gene promoters reveals 2 hubs controlling light and cytokinins response during bud outgrowth.

    Science.gov (United States)

    Roman, Hanaé; Girault, Tiffanie; Le Gourrierec, José; Leduc, Nathalie

    2017-02-01

    Bud outgrowth is under the intricate control of environmental and endogenous factors. In a recent paper, 1 we demonstrated that light perceived by Rosa buds triggers cytokinins (CK) synthesis within 3 hours in the adjacent node followed by their transport to the bud. There, CK control expression of a set of major genes (strigolactones-, auxin-, sugar sink strength-, cells division and elongation-related genes) leading to bud outgrowth in light. Conversely, under dark condition, CK accumulation and transport to the bud are repressed and no bud outgrowth occurs. In this paper, we show that the 3 expansin genes RhEXPA1,2,3 are under the control of both light and CK during bud outgrowth. In silico analysis of promoter sequences highlights 2 regions enriched in light and CK cis-regulatory elements as well as a specific cis-element in pRhEXPA3, potentially responsible for the expression patterns observed in response to CK and light.

  14. Tumor budding and E-cadherin expression are useful predictors of nodal involvement in T1 esophageal squamous cell carcinoma.

    Science.gov (United States)

    Nakagawa, Yasuo; Ohira, Masaichi; Kubo, Naoshi; Yamashita, Yoshito; Sakurai, Katsunobu; Toyokawa, Takahiro; Tanaka, Hiroaki; Muguruma, Kazuya; Shibutani, Masatsune; Yamazoe, Sadaaki; Kimura, Kenjiro; Nagahara, Hisashi; Amano, Ryosuke; Ohtani, Hiroshi; Yashiro, Masakazu; Maeda, Kiyoshi; Hirakawa, Kosei

    2013-11-01

    Endoscopic treatment has been increasingly used for T1 esophageal squamous cell carcinoma (ESCC). However, this therapy is sometimes incomplete if the depth of the T1 primary tumor reaches the muscularis mucosae or submucosal layer because these tumors have a relatively high incidence of lymph node metastasis. However, to our knowledge, no previous reports on the prediction of nodal metastasis determined by evaluating primary tumor specimens of patients with ESCC are available. A total of 55 patients with T1 ESCC invading as deep as the submucosal layer who underwent curative esophagectomy were examined. We investigated the significance of the immunohistochemical staining of Vascular endothelial growth factor-C (VEGF-C) and E-cadherin in the primary tumor and Tumor budding for prediction of nodal metastasis. Metastasis to the regional lymph nodes was observed in 26 cases (47.3%) in this setting. VEGF-C expression and reduced E-Cadherin expression in the primary tumor was observed in 32 (58.1%) and 38 cases (69.1%), respectively. High-grade tumor budding was observed in 29 cases (52.7%). E-cadherin expression and tumor budding were closely correlated with nodal metastasis (p=0.04 and tumor budding was significantly correlated (ptumor budding and E-cadherin expression for nodal metastasis were 67.3% and 65.4% respectively, comparable with the one of lymphatic involvement (63%). Tumor budding (ptumor budding are observed in primary tumor specimen.

  15. The timing of bud break in warming conditions: variation among seven sympatric conifer species from Eastern Canada.

    Science.gov (United States)

    Rossi, Sergio; Isabel, Nathalie

    2017-06-23

    Phenological changes are expected with the ongoing global warming, which could create mismatches in the growth patterns among sympatric species or create synchrony with insect herbivores. In this study, we performed a comparative assessment of the timings of bud break among seven conifer species of Eastern Canada by evaluating seedling development in growth chambers under different temperatures (16, 20 and 24 °C). Bud break occurred earliest in Larix laricina, while Pinus strobus and Pinus resinosa had the latest. Warmer conditions advanced bud break, with the greatest effects being observed at the lower temperatures. Mixed models estimated that one additional degree of temperature produced advancements of 5.3 and 2.1 days at 16 and 20 °C, respectively. The hypothesis of an asynchronous change between species under warming was demonstrated only for the last phenological phases (split buds and exposed shoots), and principally in pines. Abies balsamea showed changes in bud break comparable with the other species analysed, rejecting the hypothesis of mismatches under warmer conditions. The observed non-linear responses of the timings of bud break to warming suggest that the major changes in bud phenology should be expected at the lowest temperatures.

  16. β-catenin is required for taste bud cell renewal and behavioral taste perception in adult mice

    Science.gov (United States)

    Gaillard, Dany; Xu, Mingang; Millar, Sarah E.

    2017-01-01

    Taste stimuli are transduced by taste buds and transmitted to the brain via afferent gustatory fibers. Renewal of taste receptor cells from actively dividing progenitors is finely tuned to maintain taste sensitivity throughout life. We show that conditional β-catenin deletion in mouse taste progenitors leads to rapid depletion of progenitors and Shh+ precursors, which in turn causes taste bud loss, followed by loss of gustatory nerve fibers. In addition, our data suggest LEF1, TCF7 and Wnt3 are involved in a Wnt pathway regulatory feedback loop that controls taste cell renewal in the circumvallate papilla epithelium. Unexpectedly, taste bud decline is greater in the anterior tongue and palate than in the posterior tongue. Mutant mice with this regional pattern of taste bud loss were unable to discern sweet at any concentration, but could distinguish bitter stimuli, albeit with reduced sensitivity. Our findings are consistent with published reports wherein anterior taste buds have higher sweet sensitivity while posterior taste buds are better tuned to bitter, and suggest β-catenin plays a greater role in renewal of anterior versus posterior taste buds. PMID:28846687

  17. Release of apical dominance in potato tuber is accompanied by programmed cell death in the apical bud meristem.

    Science.gov (United States)

    Teper-Bamnolker, Paula; Buskila, Yossi; Lopesco, Yael; Ben-Dor, Shifra; Saad, Inbal; Holdengreber, Vered; Belausov, Eduard; Zemach, Hanita; Ori, Naomi; Lers, Amnon; Eshel, Dani

    2012-04-01

    Potato (Solanum tuberosum) tuber, a swollen underground stem, is used as a model system for the study of dormancy release and sprouting. Natural dormancy release, at room temperature, is initiated by tuber apical bud meristem (TAB-meristem) sprouting characterized by apical dominance (AD). Dormancy is shortened by treatments such as bromoethane (BE), which mimics the phenotype of dormancy release in cold storage by inducing early sprouting of several buds simultaneously. We studied the mechanisms governing TAB-meristem dominance release. TAB-meristem decapitation resulted in the development of increasing numbers of axillary buds with time in storage, suggesting the need for autonomous dormancy release of each bud prior to control by the apical bud. Hallmarks of programmed cell death (PCD) were identified in the TAB-meristems during normal growth, and these were more extensive when AD was lost following either extended cold storage or BE treatment. Hallmarks included DNA fragmentation, induced gene expression of vacuolar processing enzyme1 (VPE1), and elevated VPE activity. VPE1 protein was semipurified from BE-treated apical buds, and its endogenous activity was fully inhibited by a cysteinyl aspartate-specific protease-1-specific inhibitor N-Acetyl-Tyr-Val-Ala-Asp-CHO (Ac-YVAD-CHO). Transmission electron microscopy further revealed PCD-related structural alterations in the TAB-meristem of BE-treated tubers: a knob-like body in the vacuole, development of cytoplasmic vesicles, and budding-like nuclear segmentations. Treatment of tubers with BE and then VPE inhibitor induced faster growth and recovered AD in detached and nondetached apical buds, respectively. We hypothesize that PCD occurrence is associated with the weakening of tuber AD, allowing early sprouting of mature lateral buds.

  18. Sensitivity of bud burst in key tree species in the UK to recent climate variability and change

    Science.gov (United States)

    Abernethy, Rachel; Cook, Sally; Hemming, Deborah; McCarthy, Mark

    2017-04-01

    Analysing the relationship between the changing climate of the UK and the spatial and temporal distribution of spring bud burst plays an important role in understanding ecosystem functionality and predicting future phenological trends. The location and timing of bud burst of eleven species of trees alongside climatic factors such as, temperature, precipitation and hours of sunshine (photoperiod) were used to investigate: i. which species' bud burst timing experiences the greatest impact from a changing climate, ii. which climatic factor has the greatest influence on the timing of bud burst, and iii. whether the location of bud burst is influenced by climate variability. Winter heatwave duration was also analysed as part of an investigation into the relationship between temperature trends of a specific winter period and the following spring events. Geographic Information Systems (GIS) and statistical analysis tools were used to visualise spatial patterns and to analyse the phenological and climate data through regression and analysis of variance (ANOVA) tests. Where there were areas that showed a strong positive or negative relationship between phenology and climate, satellite imagery was used to calculate a Normalised Difference Vegetation Index (NDVI) and a Leaf Area Index (LAI) to further investigate the relationships found. It was expected that in the north of the UK, where bud burst tends to occur later in the year than in the south, that the bud bursts would begin to occur earlier due to increasing temperatures and increased hours of sunshine. However, initial results show that for some species, the bud burst timing tends to remain or become later in the year. Interesting results will be found when investigating the statistical significance between the changing location of the bud bursts and each climatic factor.

  19. Relationship of actin and tubulin distribution to bud growth in wild- type and morphogenetic-mutant Saccharomyces cerevisiae

    Science.gov (United States)

    1984-01-01

    The distribution of actin in wild-type cells and in morphogenetic mutants of the budding yeast Saccharomyces cerevisiae was explored by staining cells with fluorochrome-labeled phallotoxins after fixing and permeabilizing the cells by several methods. The actin appeared to be localized in a set of cortical spots or patches, as well as in a network of cytoplasmic fibers. Bundles of filaments that may possibly correspond to the fibers visualized by fluorescence were observed with the electron microscope. The putative actin spots were concentrated in small and medium-sized buds and at what were apparently the sites of incipient bud formation on unbudded cells, whereas the putative actin fibers were generally oriented along the long axes of the mother-bud pairs. In several morphogenetic mutants that form multiple, abnormally elongated buds, the actin patches were conspicuously clustered at the tips of most buds, and actin fibers were clearly oriented along the long axes of the buds. There was a strong correlation between the occurrence of active growth at particular bud tips and clustering of actin spots at those same tips. Near the end of the cell cycle in wild- type cells, actin appeared to concentrate (as a cluster of spots or a band) in the neck region connecting the mother cell to its bud. Observations made using indirect immunofluorescence with a monoclonal anti-yeast-tubulin antibody on the morphogenetic mutant cdc4 (which forms multiple, abnormally elongated buds while the nuclear cycle is arrested) revealed the surprising occurrence of multiple bundles of cytoplasmic microtubules emanating from the one duplicated spindle-pole body per cell. It seems that most or all of the buds contain one or more of these bundles of microtubules, which often can be seen to extend to the very tips of the buds. These observations are consistent with the hypotheses that actin, tubulin, or both may be involved in the polarization of growth and localization of cell-wall deposition

  20. Bud composition, branching patterns and leaf phenology in cerrado woody species.

    Science.gov (United States)

    Damascos, M A; Prado, C H B A; Ronquim, C C

    2005-11-01

    Plants have complex mechanisms of aerial biomass exposition, which depend on bud composition, the period of the year in which shoot extension occurs, branching pattern, foliage persistence, herbivory and environmental conditions. The influence of water availability and temperature on shoot growth, the bud composition, the leaf phenology, and the relationship between partial leaf fall and branching were evaluated over 3 years in Cerrado woody species Bauhinia rufa (BR), Leandra lacunosa (LL) and Miconia albicans (MA). Deciduous BR preformed organs in buds and leaves flush synchronously at the transition from the dry to the wet season. The expansion time of leaves is <1 month. Main shoots (first-order axis, A1 shoots) extended over 30 d and they did not branch. BR budding and foliage unfolds were brought about independently of inter-annual rainfall variations. By contrast, in LL and MA evergreen species, the shoot extension rate and the neoformation of aerial organs depended on rainfall. Leaf emergence was continuous for 2-6 months and lamina expansion took place over 1-4 months. The leaf life span was 5-20 months and the main A1 shoot extension happened over 122-177 d. Both evergreen species allocated biomass to shoots, leaves or flowers continuously during the year, branching in the middle of the wet season to form second-order (A2 shoots) and third-order (A3 shoots) axis in LL and A2 shoots in MA. Partial shed of A1 shoot leaves would facilitate a higher branching intensity A2 shoot production in LL than in MA. MA presented a longer leaf life span, produced a lower percentage of A2 shoots but had a higher meristem persistence on A1 and A2 shoots than LL. It was possible to identify different patterns of aerial growth in Cerrado woody species defined by shoot-linked traits such as branching pattern, bud composition, meristem persistence and leaf phenology. These related traits must be considered over and above leaf deciduousness for searching functional guilds in a

  1. Effect of Various Management Methods of Apical Flower Bud on Cut Flower Quality in Three Cultivars of Greenhouse Roses

    Directory of Open Access Journals (Sweden)

    mansour matloobi

    2017-02-01

    Full Text Available Introduction: In greenhouse roses, canopy management has been highly noted and emphasized during the past decades. It was recognized that improving canopy shape by implementing some techniques such as stem bending and flower bud removing can highly affect the marketable quality of cut roses. For most growers, the best method of flower bud treatment has not yet been described and determined physiologically. This experiment was designed to answer some questions related to this problem. Materials and Methods: A plastic commercial cut rose greenhouse was selected to carry out the trial. Three greenhouse rose cultivars, namely Eros, Cherry Brandy and Dancing Queen, were selected as the first factor, and three methods of flower bud treatment along with bending types were chosen as the second factor. Cuttings were taken from mother plants and rooted under mist conditions. The first shoot emerging from the cutting was treated at pea bud stage by one of the following methods: shoot bending at stem base with intact bud, immediate shoot bending at stem base after removing flower bud and shoot bending at stem base two weeks after flower bud removal. Some marketable stem properties including stem length, diameter and weight, and characteristics related to bud growth potential were measured, and then the data were subjected to statistical analysis. Results and Discussion: Analysis of variance showed that cultivars differ in their marketable features. Cherry Brandy produced longer cut flowers with higher stem diameter compared to the two other cultivars. This cultivar was also good in stem weight trait; however its difference from Eros was not significant. Dancing Queen did not perform well in producing high quality stems on the whole. Regarding number of days until bud release and growth, Cherry Brandy’s buds spent fewest days until growing. In many studies, the effect of cultivar on rose shoot growth quality has been documented and explained. For instance

  2. Evaluation of the Pectiform effect in the in vitro establishment of axilary buds of sweet potato clone CENSA 78-354

    Directory of Open Access Journals (Sweden)

    Orlando S. González Paneque

    2004-04-01

    Full Text Available With the purpose of studying the effect of different Pectimorf dose in the establishment and budding of axillary buds of sweet potato starting from buds, tuberous roots belonging to the clone CEMSA 78-354 were collected and placed in flasks with water in the laboratory under semicontrolled conditions to induce the sprouting of the buds. Later on, axillary buds of the buds of the tubers (establishment and axillary buds coming from in vitro plants (multiplicationwere selected as sowing material (explants, which were sowed in the Murashige and Skoog culture medium containing the MS salts, thiamin (1 mg.l-1, myoinositol (100 mg.l-1, sucrose (3%, gelryte (2 g.l-1, giberelic acid (10.0 mg.l-1 and indol-3-acetic acid (0.05 mg.l-1 and the already mentioned culture medium without the previously refered regulators of the growth with different Pectimorf concentration: 5.0, 10.0 and 15.0 mg.l-1. The morphological behavior of the in vitro plantas was evaluated to the five, fifteen and thirty days after established the buds under in vitro conditions , and different variables related with the in vitro growth of the buds were analyzed, among which we have: number of sprouted buds, height of the in vitro plants (cm, number, longitude and width of the leaves, buds with roots, longitude of the roots (cm and longitude of the petiole (cm. The best results were obtained when using the axillary buds coming from budding of tuberous roots in comparison with the axillary buds of the in vitro plants and with the use of the culture medium containing 10.0 mg.l-1 of Pectimorf. Key works: Ipomoea batatas, micropropagation, oligogalacturonides

  3. Spontaneous spheroid budding from monolayers: a potential contribution to ovarian cancer dissemination.

    Science.gov (United States)

    Pease, Jillian C; Brewer, Molly; Tirnauer, Jennifer S

    2012-07-15

    Ovarian cancer is the most lethal gynaecologic cancer, in large part because of its early dissemination and rapid development of chemotherapy resistance. Spheroids are clusters of tumor cells found in the peritoneal fluid of patients that are thought to promote this dissemination. Current models suggest that spheroids form by aggregation of single tumor cells shed from the primary tumor. Here, we demonstrate that spheroids can also form by budding directly as adherent clusters from a monolayer. Formation of budded spheroids correlated with expression of vimentin and lack of cortical E-cadherin. We also found that compared to cells grown in monolayers, cells grown as spheroids acquired progressive resistance to the chemotherapy drugs Paclitaxel and Cisplatin. This resistance could be completely reversed by dissociating the spheroids. Our observations highlight a previously unappreciated mode of spheroid formation that might have implications for tumor dissemination and chemotherapy resistance in patients, and suggest that this resistance might be reversed by spheroid dissociation.

  4. ALIX is recruited temporarily into HIV-1 budding sites at the end of gag assembly.

    Directory of Open Access Journals (Sweden)

    Pei-I Ku

    Full Text Available Polymerization of Gag on the inner leaflet of the plasma membrane drives the assembly of Human Immunodeficiency Virus 1 (HIV-1. Gag recruits components of the endosomal sorting complexes required for transport (ESCRT to facilitate membrane fission and virion release. ESCRT assembly is initiated by recruitment of ALIX and TSG101/ESCRT-I, which bind directly to the viral Gag protein and then recruit the downstream ESCRT-III and VPS4 factors to complete the budding process. In contrast to previous models, we show that ALIX is recruited transiently at the end of Gag assembly, and that most ALIX molecules are recycled into the cytosol as the virus buds, although a subset remains within the virion. Our experiments imply that ALIX is recruited to the neck of the assembling virion and is mostly recycled after virion release.

  5. ALIX is recruited temporarily into HIV-1 budding sites at the end of gag assembly.

    Science.gov (United States)

    Ku, Pei-I; Bendjennat, Mourad; Ballew, Jeff; Landesman, Michael B; Saffarian, Saveez

    2014-01-01

    Polymerization of Gag on the inner leaflet of the plasma membrane drives the assembly of Human Immunodeficiency Virus 1 (HIV-1). Gag recruits components of the endosomal sorting complexes required for transport (ESCRT) to facilitate membrane fission and virion release. ESCRT assembly is initiated by recruitment of ALIX and TSG101/ESCRT-I, which bind directly to the viral Gag protein and then recruit the downstream ESCRT-III and VPS4 factors to complete the budding process. In contrast to previous models, we show that ALIX is recruited transiently at the end of Gag assembly, and that most ALIX molecules are recycled into the cytosol as the virus buds, although a subset remains within the virion. Our experiments imply that ALIX is recruited to the neck of the assembling virion and is mostly recycled after virion release.

  6. Fine structure of kinetin-treated protonema and kinetin-induced gametophore buds in Funaria hygrometrica

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    F. Młodzianowski

    2015-01-01

    Full Text Available Besides occasional hypertrophy of grana and disintegration of stroma thylakoids occurring in some chloroplasts, no significant changes were found in ultrastructure of typical protonema cells treated for six days with kinetin. On the other hand, the fine structure of cells in kinetin--induced gametophore buds differed much from that of the protonema cells and showed characteristics of cells of with high metabolic activity and high division rates. The results indicate that cytokinins enhance development and differentiation in the protonema by activating only some of its cells, whereas the others remain unchanged or show symptoms of destruction and ageing. This is supported by the fact that in the presence of chloramphenicol, which prevents bud induction, kinetin acts synergistically with the inhibitor in producing degeneration and destruction of chloroplasts.

  7. Spontaneous spheroid budding from monolayers: a potential contribution to ovarian cancer dissemination

    Directory of Open Access Journals (Sweden)

    Jillian C. Pease

    2012-05-01

    Ovarian cancer is the most lethal gynaecologic cancer, in large part because of its early dissemination and rapid development of chemotherapy resistance. Spheroids are clusters of tumor cells found in the peritoneal fluid of patients that are thought to promote this dissemination. Current models suggest that spheroids form by aggregation of single tumor cells shed from the primary tumor. Here, we demonstrate that spheroids can also form by budding directly as adherent clusters from a monolayer. Formation of budded spheroids correlated with expression of vimentin and lack of cortical E-cadherin. We also found that compared to cells grown in monolayers, cells grown as spheroids acquired progressive resistance to the chemotherapy drugs Paclitaxel and Cisplatin. This resistance could be completely reversed by dissociating the spheroids. Our observations highlight a previously unappreciated mode of spheroid formation that might have implications for tumor dissemination and chemotherapy resistance in patients, and suggest that this resistance might be reversed by spheroid dissociation.

  8. Chromosome Segregation in Budding Yeast: Sister Chromatid Cohesion and Related Mechanisms

    Science.gov (United States)

    2014-01-01

    Studies on budding yeast have exposed the highly conserved mechanisms by which duplicated chromosomes are evenly distributed to daughter cells at the metaphase–anaphase transition. The establishment of proteinaceous bridges between sister chromatids, a function provided by a ring-shaped complex known as cohesin, is central to accurate segregation. It is the destruction of this cohesin that triggers the segregation of chromosomes following their proper attachment to microtubules. Since it is irreversible, this process must be tightly controlled and driven to completion. Furthermore, during meiosis, modifications must be put in place to allow the segregation of maternal and paternal chromosomes in the first division for gamete formation. Here, I review the pioneering work from budding yeast that has led to a molecular understanding of the establishment and destruction of cohesion. PMID:24395824

  9. Chemical and biological studies of Ribes nigrum L. buds essential oil.

    Science.gov (United States)

    Oprea, Eliza; Radulescu, Valeria; Balotescu, Carmen; Lazar, Veronica; Bucur, Marcela; Mladin, Paulina; Farcasanu, Ileana Cornelia

    2008-01-01

    Ribes nigrum buds are used in medicine for the diuretic and antiseptic properties of their volatile compounds. We present in this paper comparative data concerning the chemical composition of Ribes nigrum buds essential oils obtained from three blackcurrant varieties. Essential oils were isolated by steam distillation and were analyzed by gas chromatography coupled with mass spectrometry. The Ribes nigrum essential oils (extracted from all three varieties) exhibited similar and large antibacterial spectrum, acting against Acinetobacter (A.) baumanii, Escherichia (E.) coli, Pseudomonas (P.) aeruginoasa and Staphylococcus (S.) aureus, as proved by the very low MIC values observed for the respective strains. The subinhibitory concentrations of the essential oils induced a decrease in the bacterial ability to colonize the inert substratum for A. baumanii, E. coli and S. aureus, demonstrating that besides the bactericidal activity, the Ribes nigrum essential oils also exhibit anti-pathogenic potential.

  10. Antiviral activity of leaf-bud gum-resin of Tarenna asiatica

    OpenAIRE

    Vatsavaya Ramabharathi; Divi Venkata Ramana Saigopal; Galla Rajitha

    2014-01-01

    The leaf-bud exudate of Tarenna asiatica (Rubiaceae: Ixoroideae, Pavetteae) is investigated for its biological activity. The crude benzene extract and corymbosin (pure compound isolated) were screened for antiviral activity by using ELISA and PCR methods against animal (blue tongue and chikungunya) and plant (papaya ring spot, sesbania mosaic and common bean mosaic) viruses. Both corymbosin and benzene extract showed significant antiviral activity though corymbosin was found relatively more p...

  11. 1-MCP pretreatment prevents bud and flower abscission in Dendrobium orchids

    NARCIS (Netherlands)

    Uthaichay, N.; Ketsa, S.; Doorn, van W.G.

    2007-01-01

    Dendrobium orchid inflorescences were treated for 4 h at 25 °C with or without 100¿500 nl/l 1-MCP and were then placed in water at 25 °C to follow abscission. In controls, depending on the experiment, 20¿80% of the floral buds and 0¿20% of the open flowers abscised within 1 week. The 1-MCP

  12. Generation of micronuclei during interphase by coupling between cytoplasmic membrane blebbing and nuclear budding.

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    Koh-ichi Utani

    Full Text Available Micronucleation, mediated by interphase nuclear budding, has been repeatedly suggested, but the process is still enigmatic. In the present study, we confirmed the previous observation that there are lamin B1-negative micronuclei in addition to the positive ones. A large cytoplasmic bleb was found to frequently entrap lamin B1-negative micronuclei, which were connected to the nucleus by a thin chromatin stalk. At the bottom of the stalk, the nuclear lamin B1 structure appeared broken. Chromatin extrusion through lamina breaks has been referred to as herniation or a blister of the nucleus, and has been observed after the expression of viral proteins. A cell line in which extrachromosomal double minutes and lamin B1 protein were simultaneously visualized in different colors in live cells was established. By using these cells, time-lapse microscopy revealed that cytoplasmic membrane blebbing occurred simultaneously with the extrusion of nuclear content, which generated lamin B1-negative micronuclei during interphase. Furthermore, activation of cytoplasmic membrane blebbing by the addition of fresh serum or camptothecin induced nuclear budding within 1 to 10 minutes, which suggested that blebbing might be the cause of the budding. After the induction of blebbing, the frequency of lamin-negative micronuclei increased. The budding was most frequent during S phase and more efficiently entrapped small extrachromosomal chromatin than the large chromosome arm. Based on these results, we suggest a novel mechanism in which cytoplasmic membrane dynamics pulls the chromatin out of the nucleus through the lamina break. Evidence for such a mechanism was obtained in certain cancer cell lines including human COLO 320 and HeLa. The mechanism could significantly perturb the genome and influence cancer cell phenotypes.

  13. Asc1 supports cell-wall integrity near bud sites by a Pkc1 independent mechanism.

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    Daniel Melamed

    Full Text Available BACKGROUND: The yeast ribosomal protein Asc1 is a WD-protein family member. Its mammalian ortholog, RACK1 was initially discovered as a receptor for activated protein C kinase (PKC that functions to maintain the active conformation of PKC and to support its movement to target sites. In the budding yeast though, a connection between Asc1p and the PKC signaling pathway has never been reported. METHODOLOGY/PRINCIPAL FINDINGS: In the present study we found that asc1-deletion mutant (asc1Delta presents some of the hallmarks of PKC signaling mutants. These include an increased sensitivity to staurosporine, a specific Pkc1p inhibitor, and susceptibility to cell-wall perturbing treatments such as hypotonic- and heat shock conditions and zymolase treatment. Microscopic analysis of asc1Delta cells revealed cell-wall invaginations near bud sites after exposure to hypotonic conditions, and the dynamic of cells' survival after this stress further supports the involvement of Asc1p in maintaining the cell-wall integrity during the mid-to late stages of bud formation. Genetic interactions between asc1 and pkc1 reveal synergistic sensitivities of a double-knock out mutant (asc1Delta/pkc1Delta to cell-wall stress conditions, and high basal level of PKC signaling in asc1Delta. Furthermore, Asc1p has no effect on the cellular distribution or redistribution of Pkc1p at optimal or at cell-wall stress conditions. CONCLUSIONS/SIGNIFICANCE: Taken together, our data support the idea that unlike its mammalian orthologs, Asc1p acts remotely from Pkc1p, to regulate the integrity of the cell-wall. We speculate that its role is exerted through translation regulation of bud-site related mRNAs during cells' growth.

  14. A new hetero dimeric terpenoid derivative, japonicaside C, from the flower buds of Lonicera japonica.

    Science.gov (United States)

    Li, Hai-Bo; Yu, Yang; Mei, Yu-Dan; Meng, Zhao-Qing; Wang, Zhen-Zhong; Huang, Wen-Zhe; Xiao, Wei; Yao, Xin-Sheng

    2017-01-01

    A rare hetero dimeric terpenoid derivative, named japonicaside C, together with five known secoiridoid gloucosides were isolated from the flower buds of Lonicera japonica. The structures of these compounds were established on the basis of spectroscopic analyses. Japonicaside C is the first representative of a novel type of hetero dimeric terpenoid, biogenetically originated from a guaiane-type sesquiterpenoid and a secoiridoid glucoside. Anti-virus activity of the isolated compounds was evaluated in vitro.

  15. Calcium Regulation of Hemorrhagic Fever Virus Budding: Mechanistic Implications for Host-Oriented Therapeutic Intervention.

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    Ziying Han

    2015-10-01

    Full Text Available Hemorrhagic fever viruses, including the filoviruses (Ebola and Marburg and arenaviruses (Lassa and Junín viruses, are serious human pathogens for which there are currently no FDA approved therapeutics or vaccines. Importantly, transmission of these viruses, and specifically late steps of budding, critically depend upon host cell machinery. Consequently, strategies which target these mechanisms represent potential targets for broad spectrum host oriented therapeutics. An important cellular signal implicated previously in EBOV budding is calcium. Indeed, host cell calcium signals are increasingly being recognized to play a role in steps of entry, replication, and transmission for a range of viruses, but if and how filoviruses and arenaviruses mobilize calcium and the precise stage of virus transmission regulated by calcium have not been defined. Here we demonstrate that expression of matrix proteins from both filoviruses and arenaviruses triggers an increase in host cytoplasmic Ca2+ concentration by a mechanism that requires host Orai1 channels. Furthermore, we demonstrate that Orai1 regulates both VLP and infectious filovirus and arenavirus production and spread. Notably, suppression of the protein that triggers Orai activation (Stromal Interaction Molecule 1, STIM1 and genetic inactivation or pharmacological blockade of Orai1 channels inhibits VLP and infectious virus egress. These findings are highly significant as they expand our understanding of host mechanisms that may broadly control enveloped RNA virus budding, and they establish Orai and STIM1 as novel targets for broad-spectrum host-oriented therapeutics to combat these emerging BSL-4 pathogens and potentially other enveloped RNA viruses that bud via similar mechanisms.

  16. The budding yeast Ipl1/Aurora protein kinase regulates mitotic spindle disassembly

    OpenAIRE

    Buvelot, Stéphanie; Tatsutani, Sean Y.; Vermaak, Danielle; Biggins, Sue

    2003-01-01

    Ipl1p is the budding yeast member of the Aurora family of protein kinases, critical regulators of genomic stability that are required for chromosome segregation, the spindle checkpoint, and cytokinesis. Using time-lapse microscopy, we found that Ipl1p also has a function in mitotic spindle disassembly that is separable from its previously identified roles. Ipl1–GFP localizes to kinetochores from G1 to metaphase, transfers to the spindle after metaphase, and accumulates at the spindle midzone ...

  17. Analysis of the transcriptional responses in inflorescence buds of Jatropha curcas exposed to cytokinin treatment.

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    Chen, Mao-Sheng; Pan, Bang-Zhen; Wang, Gui-Juan; Ni, Jun; Niu, Longjian; Xu, Zeng-Fu

    2014-11-30

    Jatropha curcas L. is a potential biofuel plant. Application of exogenous cytokinin (6-benzyladenine, BA) on its inflorescence buds can significantly increase the number of female flowers, thereby improving seed yield. To investigate which genes and signal pathways are involved in the response to cytokinin in J. curcas inflorescence buds, we monitored transcriptional activity in inflorescences at 0, 3, 12, 24, and 48 h after BA treatment using a microarray. We detected 5,555 differentially expressed transcripts over the course of the experiment, which could be grouped into 12 distinct temporal expression patterns. We also identified 31 and 131 transcripts in J. curcas whose homologs in model plants function in flowering and phytohormonal signaling pathways, respectively. According to the transcriptional analysis of genes involved in flower development, we hypothesized that BA treatment delays floral organ formation by inhibiting the transcription of the A, B and E classes of floral organ-identity genes, which would allow more time to generate more floral primordia in inflorescence meristems, thereby enhancing inflorescence branching and significantly increasing flower number per inflorescence. BA treatment might also play an important role in maintaining the flowering signals by activating the transcription of GIGANTEA (GI) and inactivating the transcription of CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) and TERMINAL FLOWER 1b (TFL1b). In addition, exogenous cytokinin treatment could regulate the expression of genes involved in the metabolism and signaling of other phytohormones, indicating that cytokinin and other phytohormones jointly regulate flower development in J. curcas inflorescence buds. Our study provides a framework to better understand the molecular mechanisms underlying changes in flowering traits in response to cytokinin treatment in J. curcas inflorescence buds. The results provide valuable information related to the mechanisms of cross-talk among

  18. An analysis of chick limb bud intercellular adhesion underlying the establishment of cartilage aggregates in suspension culture.

    Science.gov (United States)

    Bee, J A; von der Mark, K

    1990-07-01

    To examine the mechanism of intercellular adhesion in the establishment of limb skeletal elements we have investigated the process of limb bud cell aggregation in vitro. Limb bud cells are aggregation-competent immediately after their trypsin:collagenase dissociation in the absence of calcium. This aggregation is largely Ca2(+)-independent (CI) and is completely and reversibly inhibited by cycloheximide. In contrast, when limb bud cells are first allowed to recover from Ca2(+)-free trypsin:collagenase dissociation, aggregation of the surviving population is exclusively Ca2(+)-dependent (CD) and completely and reversibly inhibited by cycloheximide. The presence of exogenous calcium during initial cell dissociation retains a functional CD aggregation mechanism. However, incubation of such cells with EGTA releases the CD component and converts the cells to a predominantly CI aggregation. Rabbits were immunized with limb bud cells exhibiting the recovered CD aggregation mechanism and the resulting immune sera were screened for their effect on cell aggregation. Relative to pre-immune sera, intact immune IgG agglutinated dissociated limb bud cells whilst immune Fab fragments inhibited their aggregation. The aggregation-inhibiting antiserum recognizes five major limb bud cell surface components with apparent molecular weights of 72K, 50K, 23K, 14.5K and 8.5K (K = 10(3) Mr), respectively. Limb bud cell surface plasma membranes were isolated by sucrose gradient density centrifugation and detergent-solubilized proteins coupled to Sepharose 4B with cyanogen bromide. Equivalent cell surface plasma membrane proteins were 125I-iodinated and applied to the affinity column. Limb bud cell surface protein affinity chromatography in the presence of exogenous calcium yields a single protein with an apparent molecular weight of approximately 8.5 K. This protein molecule elutes at 0.6 M NaCl, indicating a high affinity, is recognized by the aggregation-inhibiting antiserum, and is

  19. Carotenoid and tocopherol composition of leaves, buds, and flowers of Capparis spinosa grown wild in Tunisia.

    Science.gov (United States)

    Tlili, Nizar; Nasri, Nizar; Saadaoui, Ezzeddine; Khaldi, Abdelhamid; Triki, Saida

    2009-06-24

    High-performance liquid chromatography was used to determine carotenoids (beta-carotene, lutein, neoxanthin, and violaxanthin) and tocopherols of leaves, buds, and flowers of Tunisian Capparis spinosa. This plant shows strong resistance to hard environmental conditions, and it is one of the most commonly found aromatics in the Mediterranean kitchen. In this study, the means of the total carotenoids were 3452.5 +/- 1639.4, 1002 +/- 518.5, and 342.7 +/- 187.9 microg/g fresh weight (FW) in leaves, buds, and flowers, respectively. Lutein accounts for the high content. Violaxanthin provided the lowest portion of the total carotenoids. The principal form of tocopherol detected in leaves was alpha-tocopherol (20.19 +/- 10 mg/100 g FW). In buds and flowers, there were both alpha- (49.12 +/- 17.48 and 28.68 +/- 9.13 mg/100 g FW, respectively) and gamma-tocopherol (48.13 +/- 15.08 and 27.8 +/- 16.01 mg/100 g FW, respectively). The combined content of pro-vitamin A and vitamin E in capers encourages researchers to more explore and find developments for this plant.

  20. Form and function of topologically associating genomic domains in budding yeast.

    Science.gov (United States)

    Eser, Umut; Chandler-Brown, Devon; Ay, Ferhat; Straight, Aaron F; Duan, Zhijun; Noble, William Stafford; Skotheim, Jan M

    2017-04-11

    The genome of metazoan cells is organized into topologically associating domains (TADs) that have similar histone modifications, transcription level, and DNA replication timing. Although similar structures appear to be conserved in fission yeast, computational modeling and analysis of high-throughput chromosome conformation capture (Hi-C) data have been used to argue that the small, highly constrained budding yeast chromosomes could not have these structures. In contrast, herein we analyze Hi-C data for budding yeast and identify 200-kb scale TADs, whose boundaries are enriched for transcriptional activity. Furthermore, these boundaries separate regions of similarly timed replication origins connecting the long-known effect of genomic context on replication timing to genome architecture. To investigate the molecular basis of TAD formation, we performed Hi-C experiments on cells depleted for the Forkhead transcription factors, Fkh1 and Fkh2, previously associated with replication timing. Forkhead factors do not regulate TAD formation, but do promote longer-range genomic interactions and control interactions between origins near the centromere. Thus, our work defines spatial organization within the budding yeast nucleus, demonstrates the conserved role of genome architecture in regulating DNA replication, and identifies a molecular mechanism specifically regulating interactions between pericentric origins.

  1. Analysis of differential gene expression during floral bud abortion in radish (Raphanus sativus L.).

    Science.gov (United States)

    Zhang, J; Sun, X L; Zhang, L G; Hui, M X; Zhang, M K

    2013-07-24

    Radish floral bud abortion (FBA) is an adverse biological phenomenon that occurs during reproduction. Although FBA occurs frequently, its mechanism remains unknown. To elucidate the molecular mechanism underlying FBA, we detected gene expression differences between aborted and normal buds of radish using cDNA-amplified fragment length polymorphism (AFLP) and real-time polymerase chain reaction (real-time PCR). A total of 221 differentially expressed transcript-derived fragments (TDFs) were detected by 256 cDNA-AFLP primer combinations, of which 114 were upregulated and 107 were downregulated in the aborted buds. A total of 54 TDFs were cloned and sequenced. A BLAST search revealed that all TDFs have homologous sequences and 29 of these corresponded to known genes, whose functions were mainly related to metabolism, stimulus response, transcriptional regulation, and transportation. Expressions of 6 TDFs with different functions were further analyzed by real-time PCR yielding expression profiling results consistent with the cDNA-AFLP analysis. Our results indicated that radish FBA is related to abnormalities in various physiological and biochemical plant processes.

  2. Reverse engineering liver buds through self-driven condensation and organization towards medical application.

    Science.gov (United States)

    Shinozawa, Tadahiro; Yoshikawa, Hiroshi Y; Takebe, Takanori

    2016-12-15

    The self-organizing tissue-based approach coupled with induced pluripotent stem (iPS) cell technology is evolving as a promising field for designing organoids in culture and is expected to achieve valuable practical outcomes in regenerative medicine and drug development. Organoids show properties of functional organs and represent an alternative to cell models in conventional two-dimensional differentiation platforms; moreover, organoids can be used to investigate mechanisms of development and disease, drug discovery and toxicity assessment. Towards a more complex and advanced organoid model, it is essential to incorporate multiple cell lineages including developing vessels. Using a self-condensation method, we recently demonstrated self-organizing "organ buds" of diverse systems together with human mesenchymal and endothelial progenitors, proposing a new reverse engineering method to generate a more complex organoid structure. In this section, we review characters of organ bud technology based on two important principles: self-condensation and self-organization focusing on liver bud as an example, and discuss their practicality in regenerative medicine and potential as research tools for developmental biology and drug discovery. Copyright © 2016. Published by Elsevier Inc.

  3. Nutrition induced pleomorphism and budding mode of reproduction in Deinococcus radiodurans

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    Toleti Rao S

    2009-07-01

    Full Text Available Abstract Background Morphological adaptation is an important biological function of a microorganism to cope with its environment. Pleomorphism (to exist in a number of morphological forms took centre stage in many discussions wherein a bacterium exhibits morphological transition and altered mode of reproduction in response to an environmental condition. Findings To strengthen the concept on pleomorphism in bacteria, we report here different cell morphologies of Deinococcus radiodurans in response to variation in nutrient concentration. From our studies we attempt primary evidence towards the presence of significant population of monomer cells of D. radiodurans in specific culture condition. In this report we also illustrate with scanning electron micrographs an unusual budding mode of reproduction in D. radiodurans which was not reported till date for this group of bacteria. Conclusion In a holistic view the study reflects on bacterial shape (morphotypes and the physiological adaptation to a particular nutrient environment. The discovery of budding mode of reproduction in Deinococcus will be of interest to microbiologists. It can serve as a model system to understand the mechanism of budding process at molecular level.

  4. FGF signaling is required for anterior but not posterior specification of the murine liver bud

    Science.gov (United States)

    Wang, Jikui; Rhee, Siyeon; Palaria, Amrita; Tremblay, Kimberly D.

    2014-01-01

    Background The definitive endoderm arises as a naive epithelial sheet that produces the entire gut tube and associated organs including the liver, pancreas and lungs. Murine explant studies demonstrate that Fibroblast Growth Factor (FGF) signaling from adjacent tissues is required to induce hepatic gene expression from isolated foregut endoderm. The requirement of FGF signaling during liver development is examined via small molecule inhibition during whole embryo culture. Results Loss of FGF signaling prior to hepatic induction results in morphological defects and gene expression changes that are confined to the anterior liver bud. In contrast the posterior portion of the liver bud remains relatively unaffected. Because FGF is thought to act as a morphogen during endoderm organogenesis, the ventral pancreas was also examined after FGF inhibition. Although the size of the ventral pancreas is not affected, loss of FGF signaling results in a significantly higher density of ventral pancreas cells. Conclusions The requirement for FGF-mediated induction of hepatic gene expression differs across the anterior-posterior axis of the developing liver bud. These results underscore the importance of studying tissue differentiation in the context of the whole embryo. PMID:25302779

  5. Inducing the liver: understanding the signals that promote murine liver budding.

    Science.gov (United States)

    Tremblay, Kimberly D

    2011-07-01

    The endoderm emerges as an epithelial sheet that covers the surface of the developing murine embryo. This tissue will produce the entire gut tube as well as associated digestive and respiratory organs including the thyroid, thymus, lung, liver, and pancreas. The emergence of each endodermal organ occurs in a temporally distinct manner that is dependant upon reciprocal inductive interactions between the endoderm and the underlying mesoderm. The emergence of the hepatic endoderm, which occurs using a morphological process termed liver budding, initiates during early somitogenesis in the mouse at approximately 8.25 days post-coitum (dpc). Explant and transplant studies performed in chicken and mouse have demonstrated that secreted signals from adjacent mesodermal tissues initiate the hepatic gene program from ventral-fated endoderm. Here, we review the data in support of the roles of members of the fibroblast growth factor (FGF), bone morphogenetic protein (BMP), and Wnt signaling pathways in liver budding and discover that little is known about the precise endogenous signals involved in the molecular and morphological induction of liver budding in the mouse. Copyright © 2010 Wiley-Liss, Inc.

  6. Apical constriction initiates new bud formation during monopodial branching of the embryonic chicken lung.

    Science.gov (United States)

    Kim, Hye Young; Varner, Victor D; Nelson, Celeste M

    2013-08-01

    Branching morphogenesis sculpts the airway epithelium of the lung into a tree-like structure to conduct air and promote gas exchange after birth. In the avian lung, a series of buds emerges from the dorsal surface of the primary bronchus via monopodial branching to form the conducting airways; anatomically, these buds are similar to those formed by domain branching in the mammalian lung. Here, we show that monopodial branching is initiated by apical constriction of the airway epithelium, and not by differential cell proliferation, using computational modeling and quantitative imaging of embryonic chicken lung explants. Both filamentous actin and phosphorylated myosin light chain were enriched at the apical surface of the airway epithelium during monopodial branching. Consistently, inhibiting actomyosin contractility prevented apical constriction and blocked branch initiation. Although cell proliferation was enhanced along the dorsal and ventral aspects of the primary bronchus, especially before branch formation, inhibiting proliferation had no effect on the initiation of branches. To test whether the physical forces from apical constriction alone are sufficient to drive the formation of new buds, we constructed a nonlinear, three-dimensional finite element model of the airway epithelium and used it to simulate apical constriction and proliferation in the primary bronchus. Our results suggest that, consistent with the experimental results, apical constriction is sufficient to drive the early stages of monopodial branching whereas cell proliferation is dispensable. We propose that initial folding of the airway epithelium is driven primarily by apical constriction during monopodial branching of the avian lung.

  7. The Malleable Nature of the Budding Yeast Nuclear Envelope: Flares, Fusion, and Fenestrations.

    Science.gov (United States)

    Meseroll, Rebecca A; Cohen-Fix, Orna

    2016-11-01

    In eukaryotes, the nuclear envelope (NE) physically separates nuclear components and activities from rest of the cell. The NE also provides rigidity to the nucleus and contributes to chromosome organization. At the same time, the NE is highly dynamic; it must change shape and rearrange its components during development and throughout the cell cycle, and its morphology can be altered in response to mutation and disease. Here we focus on the NE of budding yeast, Saccharomyces cerevisiae, which has several unique features: it remains intact throughout the cell cycle, expands symmetrically during interphase, elongates during mitosis and, expands asymmetrically during mitotic delay. Moreover, its NE is safely breached during mating and when large structures, such as nuclear pore complexes and the spindle pole body, are embedded into its double membrane. The budding yeast NE lacks lamins and yet the nucleus is capable of maintaining a spherical shape throughout interphase. Despite these eccentricities, studies of the budding yeast NE have uncovered interesting, and likely conserved, processes that contribute to NE dynamics. In particular, we discuss the processes that drive and enable NE expansion and the dramatic changes in the NE that lead to extensions and fenestrations. J. Cell. Physiol. 231: 2353-2360, 2016. Published 2016. This article is a U.S. Government work and is in the public domain in the USA. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  8. Evaluation of the budding and depth of invasion (BD) model in oral tongue cancer biopsies.

    Science.gov (United States)

    Almangush, Alhadi; Leivo, Ilmo; Siponen, Maria; Sundquist, Elias; Mroueh, Rayan; Mäkitie, Antti A; Soini, Ylermi; Haglund, Caj; Nieminen, Pentti; Salo, Tuula

    2017-08-02

    It is of great clinical importance to identify simple prognostic markers from preoperative biopsies that could guide treatment planning. Here, we compared tumor budding (B), depth of invasion (D), and the combined scores (i.e., budding and depth of invasion (BD) histopathologic model) in preoperative biopsies and the corresponding postoperative specimens of oral tongue squamous cell carcinoma (OTSCC). Tumor budding and depth of invasion were evaluated in the pre- and postoperative samples from 100 patients treated for OTSCC. Sensitivity and specificity statistics were used. Our results showed statistically significant (P < 0.001) relationship between pre- and postoperative BD scores. There was an agreement between the pre- and postoperative BD model scores in 83 cases (83%) with 57.1% sensitivity (95% CI 39.4 to 73.7%) and 96.9% specificity (95% CI 89.3 to 99.6%). Our findings suggest that the BD model, analyzed from representative biopsies, could be used for the treatment planning of OTSCC.

  9. Structural and Biochemical Studies of ALIX/AlP1 and Its Role in Retrovirus Budding

    Energy Technology Data Exchange (ETDEWEB)

    Fisher,R.; Chung, H.; Zhai, Q.; Robinson, H.; Sundquist, W.; Hill, C.

    2007-01-01

    ALIX/AIP1 functions in enveloped virus budding, endosomal protein sorting, and many other cellular processes. Retroviruses, including HIV-1, SIV, and EIAV, bind and recruit ALIX through YPXnL late-domain motifs (X = any residue; n = 1-3). Crystal structures reveal that human ALIX is composed of an N-terminal Bro1 domain and a central domain that is composed of two extended three-helix bundles that form elongated arms that fold back into a 'V.'. The structures also reveal conformational flexibility in the arms that suggests that the V domain may act as a flexible hinge in response to ligand binding. YPXnL late domains bind in a conserved hydrophobic pocket on the second arm near the apex of the V, whereas CHMP4/ESCRT-III proteins bind a conserved hydrophobic patch on the Bro1 domain, and both interactions are required for virus budding. ALIX therefore serves as a flexible, extended scaffold that connects retroviral Gag proteins to ESCRT-III and other cellular-budding machinery.

  10. Volatile constituents and key odorants in leaves, buds, flowers, and fruits of Laurus nobilis L.

    Science.gov (United States)

    Kilic, Ayben; Hafizoglu, Harzemsah; Kollmannsberger, Hubert; Nitz, Siegfried

    2004-03-24

    The volatiles of fresh leaves, buds, flowers, and fruits from bay (Laurus nolilis L.) were isolated by solvent extraction and analyzed by capillary gas chromatography-mass spectrometry. Their odor quality was characterized by gas chomatography-olfactometry-mass spectrometry (HRGC-O-MS) and aroma extract dilution analysis (AEDA). In fresh bay leaves 1,8-cineole was the major component, together with alpha-terpinyl acetate, sabinene, alpha-pinene, beta-pinene, beta-elemene, alpha-terpineol, linalool, and eugenol. Besides 1,8-cineole and the pinenes, the main components in flowers were alpha-eudesmol, beta-elemene, and beta-caryophyllene, in fruits (E)-beta-ocimene and biclyclogermacrene, and in buds (E)-beta-ocimene and germacrene D. The aliphatic ocimenes and farnesenes were absent in leaves. By using HRGC-O-MS 21 odor compounds were identified in fresh leaves. Application of AEDA revealed (Z)-3-hexenal (fresh green), 1,8-cineole (eucalyptus), linalool (flowery), eugenol (clove), (E)-isoeugenol (flowery), and an unidentified compound (black pepper) with the highest flavor dilution factors. Differences between buds, flowers, fruits, and leaves with regard to the identified odor compounds are presented.

  11. Apical and basal epitheliomuscular F-actin dynamics during Hydra bud evagination

    Science.gov (United States)

    Aufschnaiter, Roland; Wedlich-Söldner, Roland; Zhang, Xiaoming

    2017-01-01

    ABSTRACT Bending of 2D cell sheets is a fundamental morphogenetic mechanism during animal development and reproduction. A critical player driving cell shape during tissue bending is the actin cytoskeleton. Much of our current knowledge about actin dynamics in whole organisms stems from studies of embryonic development in bilaterian model organisms. Here, we have analyzed actin-based processes during asexual bud evagination in the simple metazoan Hydra. We created transgenic Hydra strains stably expressing the actin marker Lifeact-GFP in either ectodermal or endodermal epitheliomuscular cells. We then combined live imaging with conventional phalloidin staining to directly follow actin reorganization. Bending of the Hydra epithelial double layer is initiated by a group of epitheliomuscular cells in the endodermal layer. These cells shorten their apical-basal axis and arrange their basal muscle processes in a circular configuration. We propose that this rearrangement generates the initial forces to bend the endoderm towards the ectoderm. Convergent tissue movement in both epithelial layers towards the centre of evagination then leads to elongation and extension of the bud along its new body axis. Tissue movement into the bud is associated with lateral intercalation of epithelial cells, remodelling of apical septate junctions, and rearrangement of basal muscle processes. The work presented here extends the analysis of morphogenetic mechanisms beyond embryonic tissues of model bilaterians. PMID:28630355

  12. Freezing behaviours in wintering Cornus florida flower bud tissues revisited using MRI.

    Science.gov (United States)

    Ishikawa, Masaya; Ide, Hiroyuki; Yamazaki, Hideyuki; Murakawa, Hiroki; Kuchitsu, Kazuyuki; Price, William S; Arata, Yoji

    2016-12-01

    How plant tissues control their water behaviours (phase and movement) under subfreezing temperatures through adaptative strategies (freezing behaviours) is important for their survival. However, the fine details of freezing behaviours in complex organs and their regulation mechanisms are poorly understood, and non-invasive visualization/analysis is required. The localization/density of unfrozen water in wintering Cornus florida flower buds at subfreezing temperatures was visualized with high-resolution magnetic resonance imaging (MRI). This allowed tissue-specific freezing behaviours to be determined. MRI images revealed that individual anthers and ovules remained stably supercooled to -14 to -21 °C or lower. The signal from other floral tissues decreased during cooling to -7 °C, which likely indicates their extracellular freezing. Microscopic observation and differential thermal analyses revealed that the abrupt breakdown of supercooled individual ovules and anthers resulted in their all-or-nothing type of injuries. The distribution of ice nucleation activity in flower buds determined using a test tube-based assay corroborated which tissues primarily froze. MRI is a powerful tool for non-invasively visualizing unfrozen tissues. Freezing events and/or dehydration events can be located by digital comparison of MRI images acquired at different temperatures. Only anthers and ovules preferentially remaining unfrozen are a novel freezing behaviour in flower buds. Physicochemical and biological mechanisms/implications are discussed. © 2016 John Wiley & Sons Ltd.

  13. Early Growth Improvement Of Sugarcane Bud Ps-881 Through Cattle Breeding Liquid Waste

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    Bambang Gunawan

    2017-10-01

    Full Text Available The advantage of cattle breeding in addition to giving a positive impact providing main products such as meat and milk also have the another advance. As matter of fact the livestock business must produce waste. Within use appropriate technology by utilizing cattle liquid waste into fertilizer material after going through the processing or fermentation process to become an organic fertilizer that is beneficial to the plant. This study proposed to carry out the effect of POC concentration of cattle urine on the germination rate of sugarcane bud that is more effective and efficient. The experiment was conducted at Experimental Garden of Agricultural Faculty at the University of Merdeka Surabaya with an elevation of 5 meters above sea level. This research used Randomized Block Design RAK with 1 treatment factor. The POC concentration of cattle urine P consisted of 8 treatment levels ie 0 ml 10 ml 20 ml 30 ml 40 ml 50 ml 60 ml and 70 ml POC per liter of water. This research revealed two results first the significant influence of POC concentration of cow urine on increasing growth of sugarcane bud plant on the variables studied including bud length root number and dry weight per plant during germination period of sugarcane plant. Second The highest value achieved by treatment P7 is the concentration of 70 ml of POC of cow urine per liter of water at all observation parameters.

  14. The flipflop orphan genes are required for limb bud eversion in the Tribolium embryo

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    Susanne Thümecke

    2017-10-01

    Full Text Available Abstract Background Unlike Drosophila but similar to other arthropod and vertebrate embryos, the flour beetle Tribolium castaneum develops everted limb buds during embryogenesis. However, the molecular processes directing the evagination of epithelia are only poorly understood. Results Here we show that the newly discovered genes Tc-flipflop1 and Tc-flipflop2 are involved in regulating the directional budding of appendages. RNAi-knockdown of Tc-flipflop results in a variety of phenotypic traits. Most prominently, embryonic limb buds frequently grow inwards rather than out, leading to the development of inverted appendages inside the larval body. Moreover, affected embryos display dorsal closure defects. The Tc-flipflop genes are evolutionarily non-conserved, and their molecular function is not evident. We further found that Tc-RhoGEF2, a highly-conserved gene known to be involved in actomyosin-dependent cell movement and cell shape changes, shows a Tc-flipflop-like RNAi-phenotype. Conclusions The similarity of the inverted appendage phenotype in both the flipflop- and the RhoGEF2 RNAi gene knockdown led us to conclude that the Tc-flipflop orphan genes act in a Rho-dependent pathway that is essential for the early morphogenesis of polarised epithelial movements. Our work describes one of the few examples of an orphan gene playing a crucial role in an important developmental process.

  15. Bone morphogenetic protein 4 regulates the budding site and elongation of the mouse ureter

    Science.gov (United States)

    Miyazaki, Yoichi; Oshima, Keisuke; Fogo, Agnes; Hogan, Brigid L.M.; Ichikawa, Iekuni

    2000-01-01

    In the normal mouse embryo, Bmp4 is expressed in mesenchymal cells surrounding the Wolffian duct (WD) and ureter stalk, whereas bone morphogenetic protein (BMP) type I receptor genes are transcribed either ubiquitously (Alk3) or exclusively in the WD and ureter epithelium (Alk6). Bmp4 heterozygous null mutant mice display, with high penetrance, abnormalities that mimic human congenital anomalies of the kidney and urinary tract (CAKUT), including hypo/dysplastic kidneys, hydroureter, ectopic ureterovesical (UV) junction, and double collecting system. Analysis of mutant embryos suggests that the kidney hypo/dysplasia results from reduced branching of the ureter, whereas the ectopic UV junction and double collecting system are due to ectopic ureteral budding from the WD and accessory budding from the main ureter, respectively. In the cultured metanephros deprived of sulfated glycosaminoglycans (S-GAGs), BMP4-loaded beads partially rescue growth and elongation of the ureter. By contrast, when S-GAGs synthesis is not inhibited, BMP4 beads inhibit ureter branching and expression of Wnt 11, a target of glial cell-derived neurotrophic factor signaling. Thus, Bmp4 has 2 functions in the early morphogenesis of the kidney and urinary tract. One is to inhibit ectopic budding from the WD or the ureter stalk by antagonizing inductive signals from the metanephric mesenchyme to the illegitimate sites on the WD. The other is to promote the elongation of the branching ureter within the metanephros, thereby promoting kidney morphogenesis. PMID:10749566

  16. Role of Intra- and Peritumoral Budding in the Interdisciplinary Management of Rectal Cancer Patients

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    Inti Zlobec

    2012-01-01

    Full Text Available The presence of tumor budding (TuB at the invasive front of rectal cancers is a valuable indicator of tumor aggressiveness. Tumor buds, typically identified as single cells or small tumor cell clusters detached from the main tumor body, are characterized by loss of cell adhesion, increased migratory, and invasion potential and have been referred to as malignant stem cells. The adverse clinical outcome of patients with a high-grade TuB phenotype has consistently been demonstrated. TuB is a category IIB prognostic factor; it has yet to be investigated in the prospective setting. The value of TuB in oncological and pathological practice goes beyond its use as a simple histomorphological marker of tumor aggressiveness. In this paper, we outline three situations in which the assessment of TuB may have direct implications on treatment within the multidisciplinary management of patients with rectal cancer: (a patients with TNM stage II (i.e., T3/T4, N0 disease potentially benefitting from adjuvant therapy, (b patients with early submucosally invasive (T1, sm1-sm3 carcinomas at a high risk of nodal positivity and (c the role of intratumoral budding assessed in preoperative biopsies as a marker for lymph node and distant metastasis thus potentially aiding the identification of patients suitable for neoadjuvant therapy.

  17. Correlation of number of tumor buds and tumor stage in large bowel carcinomas

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    Đerković Branislav

    2016-01-01

    Full Text Available Standardized staging of tumors takes into account the depth of invasion of the intestinal wall and the presence of local or distant metastases, specifically focusing to precisely estimate length of patient survival. This assessment system does not fully reflect the biological behaviour of cancer individually, ie. tumor aggressiveness and ability of recurrence tumor after medical treatment. Furthermore, cancer at some patients have more aggressive growth than other carcinomas in the same clinical stage, because there are other parameters that determine the biological behaviors of colon cancer, which are not included in the standard classification of determining tumor stage. One of the recent arguments which are due in the spotlight is 'tumor budding', which represents one cell or group of up to five non-differentiated tumor cells, which are found in the stroma out of the invasive front line of cancer. There are 92 colon cancer and upper rectum processed, which are collected at General Hospital in Trebinje and Medical Center in Kosovska Mitrovica. The aim is to determine whether there is a correlation between the number of tumor budding and stage of tumors in colorectal cancer. The tumor stage is determined by Astler Coller classification. Investigation, based on x2-test, leads to the conclusion that there is not a statistical significance in tumor budding distribution in relation to tumor stage according to the Astler Coller classification (p = 0.383; p> 0.05.

  18. Chilling-responsive DEMETER-LIKE DNA demethylase mediates in poplar bud break.

    Science.gov (United States)

    Conde, Daniel; Le Gac, Anne-Laure; Perales, Mariano; Dervinis, Christopher; Kirst, Matias; Maury, Stéphane; González-Melendi, Pablo; Allona, Isabel

    2017-10-01

    Annual dormancy-growth cycle is a developmental and physiological process essential for the survival of deciduous trees in temperate and boreal forests. Seasonal control of shoot growth in woody perennials requires specific genetic programmes responding to environmental signals. The environmental-controlled mechanisms that regulate the shift between winter dormancy and the growth-promoting genetic programmes are still unknown. Here, we show that dynamics in genomic DNA methylation levels are involved in the regulation of dormancy-growth cycle in poplar. The reactivation of growth in the apical shoot during bud break process in spring is preceded by a progressive reduction of genomic DNA methylation in apex tissue. The induction in apex tissue of a chilling-dependent poplar DEMETER-LIKE 10 (PtaDML10) DNA demethylase precedes shoot growth reactivation. Transgenic poplars showing downregulation of PtaDML8/10 caused delayed bud break. Genome-wide transcriptome and methylome analysis and data mining revealed that the gene targets of DEMETER-LIKE-dependent DNA demethylation are genetically associated with bud break. These data point to a chilling-dependent DEMETER-like DNA demethylase mechanisms being involved in the shift from winter dormancy to a condition that precedes shoot apical vegetative growth in poplar. © 2017 John Wiley & Sons Ltd.

  19. A nutrient dependant switch explains mutually exclusive existence of meiosis and mitosis initiation in budding yeast.

    Science.gov (United States)

    Wannige, C T; Kulasiri, D; Samarasinghe, S

    2014-01-21

    Nutrients from living environment are vital for the survival and growth of any organism. Budding yeast diploid cells decide to grow by mitosis type cell division or decide to create unique, stress resistant spores by meiosis type cell division depending on the available nutrient conditions. To gain a molecular systems level understanding of the nutrient dependant switching between meiosis and mitosis initiation in diploid cells of budding yeast, we develop a theoretical model based on ordinary differential equations (ODEs) including the mitosis initiator and its relations to budding yeast meiosis initiation network. Our model accurately and qualitatively predicts the experimentally revealed temporal variations of related proteins under different nutrient conditions as well as the diverse mutant studies related to meiosis and mitosis initiation. Using this model, we show how the meiosis and mitosis initiators form an all-or-none type bistable switch in response to available nutrient level (mainly nitrogen). The transitions to and from meiosis or mitosis initiation states occur via saddle node bifurcation. This bidirectional switch helps the optimal usage of available nutrients and explains the mutually exclusive existence of meiosis and mitosis pathways. © 2013 Elsevier Ltd. All rights reserved.

  20. Longitudinal analysis of calorie restriction on rat taste bud morphology and expression of sweet taste modulators.

    Science.gov (United States)

    Cai, Huan; Daimon, Caitlin M; Cong, Wei-Na; Wang, Rui; Chirdon, Patrick; de Cabo, Rafael; Sévigny, Jean; Maudsley, Stuart; Martin, Bronwen

    2014-05-01

    Calorie restriction (CR) is a lifestyle intervention employed to reduce body weight and improve metabolic functions primarily via reduction of ingested carbohydrates and fats. Taste perception is highly related to functional metabolic status and body adiposity. We have previously shown that sweet taste perception diminishes with age; however, relatively little is known about the effects of various lengths of CR upon taste cell morphology and function. We investigated the effects of CR on taste bud morphology and expression of sweet taste-related modulators in 5-, 17-, and 30-month-old rats. In ad libitum (AL) and CR rats, we consistently found the following parameters altered significantly with advancing age: reduction of taste bud size and taste cell numbers per taste bud and reduced expression of sonic hedgehog, type 1 taste receptor 3 (T1r3), α-gustducin, and glucagon-like peptide-1 (GLP-1). In the oldest rats, CR affected a significant reduction of tongue T1r3, GLP-1, and α-gustducin expression compared with age-matched AL rats. Leptin receptor immunopositive cells were elevated in 17- and 30-month-old CR rats compared with age-matched AL rats. These alterations of sweet taste-related modulators, specifically during advanced aging, suggest that sweet taste perception may be altered in response to different lengths of CR.

  1. In vitro propagation of Caesalpinia spinosa (Mol. O. Kuntz from axillary buds of selected trees

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    Jenny E Núñez Núñez

    2017-07-01

    Full Text Available Guarango or tara [Caesalpinia spinosa (Mol. O. Kuntz] is a tree native to the Andes, with great economic importance and for reforestation programs. The aim of this work was to in vitro propagate this specie from axillary buds of selected trees. During in vitro establishment, the effect of sodium hypochlorite (3.0% with different times of disinfection (5.0, 10, 15 min, as well as the effect of 6-BAP on the in vitro response of buds were studied. For multiplication, different combination of 6-BAP with 0.1 mg l-1 ANA were tested. A free-growth regulator culture medium was used for rooting. The best results for in vitro establishment were achieved with a disinfection treatment with sodium hypochlorite 3.0% for 10 minutes and cultivation in a culture medium with 0.25 mg l-1 6-BAP, which 90% of buds in vitro established, with a length of 6.71 cm. The highest multiplication rate of shoot (2.88 per explant was obtained with 1.0 mg l-1 6-BAP and 0.1 mg l-1 ANA, after 60 days of culture. The 55% of these shoots developed roots in a half-strength basal salts MS culture medium free of regulators of growth.   Keywords: biodiversity, conservation, forest plant, guarango, tissue culture

  2. Molecular Evidence for Catechin Synthesis and Accumulation in Tea Buds (Camellia sinensis).

    Science.gov (United States)

    Wang, Xinzhen; Guo, Lina; Gao, Liping; Shi, Xingxing; Zhao, Xuecheng; Ma, Xiubing; Xia, Tao; Wang, Yunsheng

    2018-01-10

    Early spring buds of the Camellia sinensis variety Shuchazao were separated into two parts, including the shoot tip (ST) and non-expanded young leaves (YL), in which the synthesis and accumulation of catechins in the two parts were assessed by high-performance liquid chromatography (HPLC), p-dimethylaminocinnamaldehyde (DMACA) staining, quantitative real-time polymerase chain reaction (qRT-PCR), and in situ hybridization. HPLC showed that (-)-epigallocatechin-3-gallate (EGCG) and (-)-epicatechin-3-gallate (ECG) amounts in YL were increased significantly by 74.0 and 71.8%, respectively. The results of DMACA staining indicated that catechins in buds accumulated mainly in mesophyll cells and the bud shaft of YL. Meanwhile, qRT-PCR demonstrated that the relative expression levels of genes related to flavonoid metabolism, including CsPAL1, CsC4H1, CsC4H2, CsCHS2, CsF3'5'H1, CsDFR1, CsDFR2, and CsANR1, were significantly higher in YL than in the ST. In situ hybridization revealed that CsDFR1, CsDFR2, CsLAR, and CsANR1 were expressed in leaf primordia and YL but not in the apical meristem. These findings highlight the synthesis and accumulation patterns of catechins in different parts of the ST in C. sinensis, providing a theoretical basis for the assessment of synthesis, accumulation, and transfer patterns of catechins in tea plants.

  3. FGF signaling is required for anterior but not posterior specification of the murine liver bud.

    Science.gov (United States)

    Wang, Jikui; Rhee, Siyeon; Palaria, Amrita; Tremblay, Kimberly D

    2015-03-01

    The definitive endoderm arises as a naive epithelial sheet that produces the entire gut tube and associated organs including the liver, pancreas and lungs. Murine explant studies demonstrate that fibroblast growth factor (FGF) signaling from adjacent tissues is required to induce hepatic gene expression from isolated foregut endoderm. The requirement of FGF signaling during liver development is examined by means of small molecule inhibition during whole embryo culture. Loss of FGF signaling before hepatic induction results in morphological defects and gene expression changes that are confined to the anterior liver bud. In contrast the posterior portion of the liver bud remains relatively unaffected. Because FGF is thought to act as a morphogen during endoderm organogenesis, the ventral pancreas was also examined after FGF inhibition. Although the size of the ventral pancreas is not affected, loss of FGF signaling results in a significantly higher density of ventral pancreas cells. The requirement for FGF-mediated induction of hepatic gene expression differs across the anterior/posterior axis of the developing liver bud. These results underscore the importance of studying tissue differentiation in the context of the whole embryo. © 2014 Wiley Periodicals, Inc.

  4. Malformation of gynoecia impedes fertilisation in bud-flowering Calluna vulgaris.

    Science.gov (United States)

    Behrend, A; Borchert, T; Müller, A; Tänzer, J; Hohe, A

    2013-01-01

    In Calluna vulgaris, a common bedding plant during autumn in the northern hemisphere, the bud-blooming mutation of flower morphology is of high economic importance. Breeding of new bud-blooming cultivars suffers from poor seed set in some of the desirable bud-flowering crossing partners. In the current study, fertilisation and seed development in genotypes with good or poor seed set were monitored in detail in order to examine pre- and post-zygotic cross breeding incompatibilities. Whereas no distinct differences were detected in seed development, pollen tube growth was impeded in the pistils of genotypes characterised by poor seed set. Detailed microscopic analysis revealed malformations of the gynoecia due to imperfect fusion of carpels. Hence, a pre-zygotic mechanism hindering pollen tube growth due to malformation of gynoecia was deduced. An interaction of putative candidate genes involved in malformation of gynoecia with floral organ identity genes controlling the flower architecture is discussed. © 2012 German Botanical Society and The Royal Botanical Society of the Netherlands.

  5. SUBSTRATES UTILIZATION TO ASSESS ROOTEDNESS CAPACITY AND VIABILITY BUDS AT SOME GRAPE VARIETIES

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    Gheorghe Cristian Popescu

    2013-12-01

    Full Text Available The cultivated grapevine (Vitis vinifera L. is a fruit crop of enormous economic importance with over eight million hectares planted in vineyards worldwide. Table grapes and wines represent a considerable share of the economy in many grape and wine-producing countries. During the dormant, due to low temperatures and how to prepare grape for entrance in winter time, wood annual increases and buds may be adversely affected. The way how the vines passed by dormant period can affect the buds and wood viability and rooting ability of vine cuttings. In this study were tested on different culture substrates vine cuttings belonging to a noble variety and a hybrid vines: Merlot and Isabella. Noble grapes are a term used to describe the international variety of grapes that are most recognizable for the top quality wine they produce. In this paper was determinate total dry matter of vine cuttings, humidity of biological material, vine cuttings rooting capacity and viability status buds cuttings placed on three nutritional substrates.

  6. Pore formation on proliferating yeast Saccharomyces cerevisiae cell buds by HM-1 killer toxin.

    Science.gov (United States)

    Komiyama, T; Ohta, T; Urakami, H; Shiratori, Y; Takasuka, T; Satoh, M; Watanabe, T; Furuichi, Y

    1996-04-01

    The cytocidal effect of HM-1 produced by Hansenula mrakii on yeast Saccharomyces cerevisiae cells was studied. The HM-1 strongly inhibited the growth of S. cerevisiae cells at a low concentration (IC50: 2.1 x 10(-8) M) by reducing the number of viable cells. The killer action of HM-1 was most efficient when cells were actively proliferating. Cells in a resting state were resistant, but they became HM-1-sensitive after about 90 min of culturing at 30 degrees C, concomitantly with the increment of budding index. In association with the reduction of viable cell number, ultraviolet light-absorbing cellular components were discharged from sensitive cells. HM-1 molecules appear to bind to susceptible cells rather loosely since cells incubated with HM-1 were able to proliferate after having been washed. By phase-contrast light microscopy and scanning electron microscopy, discharge of cell material was observed at the budding portions of HM-1-treated cells. Addition of sorbitol to make the culture medium isotonic partially reduced the cell death induced by HM-1. These results suggest that HM-1 acts on the budding region of proliferating yeast cells, resulting in pore formation, leakage of cell material and eventual cell death.

  7. Spatio-temporal relief from hypoxia and production of reactive oxygen species during bud burst in grapevine (Vitis vinifera L.)

    OpenAIRE

    Meitha, K; Konnerup, D; Colmer, TD; Considine, JA; Foyer, CH; Considine, MJ

    2015-01-01

    Background and Aims: Plants regulate cellular oxygen partial pressures (pO2), together with reduction/ oxidation (redox) state to manage rapid developmental transitions such as bud burst after a period of quiescence. However, our understanding of pO2 regulation in complex meristematic organs such as buds is incomplete, and particularly lacks spatial resolution. Methods: The gradients in pO2 from the outer scales to the primary meristem complex were measured in grapevine (Vitis vinifera L.) bu...

  8. Role of Tulipa gesneriana TEOSINTE BRANCHED1 (TgTB1) in the control of axillary bud outgrowth in bulbs.

    Science.gov (United States)

    Moreno-Pachon, Natalia M; Mutimawurugo, Marie-Chantal; Heynen, Eveline; Sergeeva, Lidiya; Benders, Anne; Blilou, Ikram; Hilhorst, Henk W M; Immink, Richard G H

    2017-12-07

    Tulip vegetative reproduction. Tulips reproduce asexually by the outgrowth of their axillary meristems located in the axil of each bulb scale. The number of axillary meristems in one bulb is low, and not all of them grow out during the yearly growth cycle of the bulb. Since the degree of axillary bud outgrowth in tulip determines the success of their vegetative propagation, this study aimed at understanding the mechanism controlling the differential axillary bud activity. We used a combined physiological and "bottom-up" molecular approach to shed light on this process and found that first two inner located buds do not seem to experience dormancy during the growth cycle, while mid-located buds enter dormancy by the end of the growing season. Dormancy was assessed by weight increase and TgTB1 expression levels, a conserved TCP transcription factor and well-known master integrator of environmental and endogenous signals influencing axillary meristem outgrowth in plants. We showed that TgTB1 expression in tulip bulbs can be modulated by sucrose, cytokinin and strigolactone, just as it has been reported for other species. However, the limited growth of mid-located buds, even when their TgTB1 expression is downregulated, points at other factors, probably physical, inhibiting their growth. We conclude that the time of axillary bud initiation determines the degree of dormancy and the sink strength of the bud. Thus, development, apical dominance, sink strength, hormonal cross-talk, expression of TgTB1 and other possibly physical but unidentified players, all converge to determine the growth capacity of tulip axillary buds.

  9. Changes in Protein Profiles of Poplar Tissues during the Induction of Bud Dormancy by Short-Day Photoperiods

    OpenAIRE

    Zoran, Jeknic; Tony H.H., Chen; Department of Horticulture, 4017 Agricultural and Life Sciences, Oregon State University; Department of Horticulture, 4017 Agricultural and Life Sciences, Oregon State University

    1999-01-01

    Vegetative bud dormancy in woody perennial plants of the temperate regions is an important adaptive strategy for withstanding low winter temperatures. We used shortday(SD) photoperiods to induce bud dormancy in poplar(Populus deltoides Bartr.ex Marsh.), and characterized changes in protein profiles duriong dormancy development. Short days alone, under warm temperatures(25℃) induced a high level of dormancy comparable to that developed naturally. Under SD conditions the amounts of acetone/tric...

  10. Tumor Budding, EMT and Cancer Stem Cells in T1-2/N0 Oral Squamous Cell Carcinomas.

    Science.gov (United States)

    Attramadal, Cecilie Gjøvaag; Kumar, Sheeba; Boysen, Morten E; Dhakal, Hari Prasad; Nesland, Jahn Marthin; Bryne, Magne

    2015-11-01

    Early oral carcinomas have a high recurrence rate despite surgery with clear margins. In an attempt to classify the risk of recurrence of oral squamous cell carcinomas, we explored the significance of tumor budding, epithelial-mesenchymal transition (EMT) and certain cancer stem cell markers (CSC). Tumor budding (single cells or clusters of ≤5 cells in the tumor front, divided into high- and low-budding tumors), EMT and CSC markers were studied in 62 immunohistochemically stained slides of T1/2N0M0 oral squamous cell carcinomas. Tissues and records of follow-up were obtained from the Oslo University Hospital, Norway. Tumor budding, EMT and CSC markers were scored and analyzed. The only significant prognostic marker was tumor budding (p=0.043). Expression of the EMT marker E-cadherin was lost from the invasive front and tended to be a prognostic factor (p=0.17), and up-regulation of vimentin in tumor cells in the invasive front was found; this indicates that EMT had occurred. CSC markers were not associated with recurrence rate in the present study. A high budding index was related to poor prognosis in patients with oral cancer. Budding was associated with EMT-like changes. CSC factors were detected but reflected differentiation rather than stemness. Scoring of buds in patients with oral cancer may help discriminate invasive tumors prone to relapse, and thus, provide an indication for adjuvant therapy. Copyright© 2015 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

  11. [Study on tissue culture and plant regeneration of the stem-tips and buds of Periploca forrestii].

    Science.gov (United States)

    Liu, Zhi-Fei; Gao, Jie; Niu, Ya-Hui; Shi, Lei; Chen, Tao-Tao

    2011-11-01

    To establish the rapid propagation systems of the stem-tips and buds of Periploca forrestii. Inserted the stem-tips and buds of Periploca forrestii into MS medium with different concentrations of 6-BA, NAA and 2.4-D and induced them growing into complete plants. The optimal culture medium for bud induction is MS + 6-BA 1.0 mg/L + NAA 0.3 mg/L and the bud induction rate can reach 86.29%. The optimal culture medium for stem-tips induction is MS +6-BA 2.0 mg/L + NAA 0.5 mg/L and the bud induction rate can reach 86.29%. The optimal culture medium for bud multiplication is MS + 6-BA 2.0 mg/L + NAA 0.1 mg/L and the multiplication coefficient can reach 2.10. The best rooting medium is 1/2 MS + IBA 0.5 mg/L and the rooting rate is 53.33%.

  12. Cytokeratin immunohistochemistry improves interobserver variability between unskilled pathologists in the evaluation of tumor budding in T1 colorectal cancer.

    Science.gov (United States)

    Kai, Keita; Aishima, Shinichi; Aoki, Shigehisa; Takase, Yukari; Uchihashi, Kazuyoshi; Masuda, Masanori; Nishijima-Matsunobu, Aki; Yamamoto, Mihoko; Ide, Kousuke; Nakayama, Atsushi; Yamasaki, Makiko; Toda, Shuji

    2016-02-01

    Tumor budding is a major risk factor for T1 colorectal cancer. Quality control of the pathological diagnosis of budding is crucial, irrespective of the pathologist's experience. This study examines the interobserver variability according to pathologists' experience and evaluates the influence of cytokeratin (CK) immunostaining in the assessment of budding. Hematoxylin-eosin (HE) and CK-immunostained slides of 40 cases with T1 primary colorectal cancer were examined. Budding grades were individually evaluated by 12 pathologists who we categorized into three groups by their experience (expert, with >10 years of experience (n = 4), senior, with 5-10 years (n = 4), and junior, budding grades compared to the less-experienced pathologists. In the junior group, the interobserver variability obtained with HE slides was poor, but it was markedly improved in the evaluation using CK-immunostained slides. The benefit of CK immunostaining was only slight in the expert group. CK immunostaining would be useful when a pathologist is not experienced enough or does not have enough confidence in the assessment of budding. © 2016 Japanese Society of Pathology and John Wiley & Sons Australia, Ltd.

  13. Tumor budding as a useful prognostic marker in T1-stage squamous cell carcinoma of the esophagus.

    Science.gov (United States)

    Teramoto, Hitoshi; Koike, Masahiko; Tanaka, Chie; Yamada, Suguru; Nakayama, Goro; Fujii, Tsutomu; Sugimoto, Hiroyuki; Fujiwara, Michitaka; Suzuki, Yasuhiko; Kodera, Yasuhiro

    2013-07-01

    Establishing a new prognostic factor for early-stage cancer may seem difficult due to the small number of disease-specific deaths. Tumor budding has been recognized as a useful microscopic finding reflecting biological activity of the tumor. Tumor budding stand for isolated single cancer cells and cell clusters scattered beyond the tumor margin at the invasive front. It was searched for in the resected esophagus with T1 squamous cell carcinoma (SCC), and the correlation between the tumor budding, patient survival, and various pathologic factors were analyzed to verify whether tumor budding is a prognostic factor in superficial esophageal cancer. Seventy-nine patients undergoing curative esophagectomy were assigned to frequent (n = 29) and rare (n = 50) groups according to the microscopically observed frequency of tumor budding in the tumor. Three-year survival rates after esophagectomy were 48.8% for the frequent group and 94.5% for the rare group. Multivariate analysis using the Cox proportional hazards model identified this morphological variable as a significant independent prognostic factor. Tumor budding reflects the biological activity of the tumor and may be a useful prognostic indicator even in early-stage SCC of esophagus. Copyright © 2013 Wiley Periodicals, Inc.

  14. Plant regeneration and floral bud formation from intact floral parts of African violet (Saintpaulia ionantha H. Wendl.) cultured in vitro.

    Science.gov (United States)

    Daud, N; Taha, R M

    2008-04-01

    Intact immature flower buds of African violet (Saintpaulia ionantha H. Wendl.) were used as explant sources for in vitro studies. The effect of exogenous hormones, NAA and BAP on the indirect organogenesis of this species was observed. Callus was formed on the cut end (base) of pedicels of floral buds where they were in contact with the medium. When maintained on the same medium, callus was differentiated into adventitious shoots after 10 weeks in culture. MS media supplemented with 2.0 mg L(-1) NAA and 1.0 mg L(-1) BAP gave the highest number of sterile or vegetative floral buds from the surface of callus of the explants, but these buds failed to develop further. The floral buds were expanded as abnormal flowers. The floral structures were smaller in size compared to intact flowers. Petals (corolla) were white to purple in colour but did not form any reproductive organs, i.e., stamens or pistils. All sterile or vegetative floral buds and abnormal flowers survived for 3 months in culture but failed to reach anthesis.

  15. Effect of bud burst forcing on transcript expression of selected genes in needles of Norway spruce during autumn.

    Science.gov (United States)

    Asante, Daniel K A; Yakovlev, Igor A; Fossdal, Carl Gunnar; Timmerhaus, Gerrit; Partanen, Jouni; Johnsen, Oystein

    2009-08-01

    Expression of selected genes in needles of Norway spruce (Picea abies [L.] Karst) was investigated by following their transcription levels during late autumn. Transcription was assessed in mature needles which likely serve as sensor of environmental cues that enable trees in the temperate and boreal regions to change between stages of growth, frost tolerance and bud dormancy. Samples were collected from grafts kept under outdoor conditions and after bud burst forcing in greenhouse at 20 degrees C (12 h darkness) for one week. Transcription was assayed with real-time RT-PCR. During the sampling period, chilling requirement was partially fulfilled, and time to bud burst after forcing was decreased. Of the 27 transcripts studied, expression of 16 was significantly affected either by forcing, sampling time, or interaction between them. PaSAP, PaACP, PaSGS3, PaWRKY, PaDIR9, PaCCCH and dehydrin genes responded drastically to forcing temperatures at all sampling points, showing no correlation with readiness for bud burst. Expression patterns of some vernalization pathway gene homologs PaVIN3, and also of PaMDC, PaLOV1 and PaDAL3 had a clear opposite trends between forcing and outdoor conditions, which could imply their role in chilling accumulation and bud burst regulation/cold acclimation. These genes could constitute putative candidates for further detailed study, whose regulation in needles may be involved in preparation towards bud burst and chilling accumulation sensing.

  16. Nutritional modulation of mouse and human liver bud growth through a branched-chain amino acid metabolism.

    Science.gov (United States)

    Koike, Hiroyuki; Zhang, Ran-Ran; Ueno, Yasuharu; Sekine, Keisuke; Zheng, Yun-Wen; Takebe, Takanori; Taniguchi, Hideki

    2017-03-15

    Liver bud progenitors experience a transient amplification during the early organ growth phase, yet the mechanism responsible is not fully understood. Collective evidence highlights the specific requirements in stem cell metabolism for expanding organ progenitors during organogenesis and regeneration. Here, transcriptome analyses show that progenitors of the mouse and human liver bud growth stage specifically express the gene branched chain aminotransferase 1, encoding a known breakdown enzyme of branched-chain amino acids (BCAAs) for energy generation. Global metabolome analysis confirmed the active consumption of BCAAs in the growing liver bud, but not in the later fetal or adult liver. Consistently, maternal dietary restriction of BCAAs during pregnancy significantly abrogated the conceptus liver bud growth capability through a striking defect in hepatic progenitor expansion. Under defined conditions, the supplementation of L-valine specifically among the BCAAs promoted rigorous growth of the human liver bud organoid in culture by selectively amplifying self-renewing bi-potent hepatic progenitor cells. These results highlight a previously underappreciated role of branched-chain amino acid metabolism in regulating mouse and human liver bud growth that can be modulated by maternal nutrition in vivo or cultural supplement in vitro. © 2017. Published by The Company of Biologists Ltd.

  17. MicroRNA-200b is downregulated in colon cancer budding cells

    Science.gov (United States)

    Lindebjerg, Jan; Nielsen, Boye Schnack; Hansen, Torben Frøstrup; Sørensen, Flemming Brandt

    2017-01-01

    Background The microRNA-200 (miR-200) family acts as a major suppressor of epithelial-mesenchymal transition (EMT). Impaired miR-200 expression may lead to EMT initiation and eventually cancer dissemination. The presence of tumor budding cells (TBC) is associated with metastasis and poor prognosis, and molecular similarities to EMT indicate that these cells may reflect ongoing EMT. The aim of this study was to investigate the expression of miR-200b in budding cells of colon cancer and the relationship with the EMT-markers E-cadherin, β-catenin and laminin-5γ2. Material & methods MiR-200b was investigated by in situ hybridization in 58 cases of stage II (n = 36) and III colon (n = 22) cancers with tumor budding. Expression of E-cadherin, β-catenin and laminin-5γ2 was examined by immunohistochemistry. A multiplex fluorescence assay combining miR-200b with cytokeratin and laminin-5γ2 was employed on a subset of 16 samples. Results MiR-200b was downregulated in the TBC at the invasive front of 41 out of 58 (71%) cases. The decline was present in both mismatch satellite stable and instable adenocarcinomas. The majority of cases also showed loss of membranous E-cadherin and increased nuclear β-catenin in the TBC, while laminin-5γ2 expression was upregulated at the invasive front and in the tumor buds of approximately half the adenocarcinomas. However, the miR-200b decline was not statistically associated with the expression of any of the EMT-markers. The miR-200b decline was also documented by multiplex fluorescence. Fourteen out of fifteen cases showed a decrease in miR-200b expression in the majority of the TBC, but no obvious relationship between miR-200b and laminin-5γ2 expression was observed. Conclusion: The findings support the assumption of a miR-200b related downregulation in colon cancer budding cells. Whether miR-200b expression may be of clinical significance awaits further studies. PMID:28552992

  18. The effect of the times and the budding methods on the quality of young trees and the nursery efficiency of cherry trees cv. 'Łutówka'

    Directory of Open Access Journals (Sweden)

    Piotr Baryła

    2012-12-01

    Full Text Available The studies concerning the effect of the times and the methods of budding on the growth of young cherry trees were conducted in the years 1997-2000 at Felin Experimental Farm of Lublin Agricultural University. The objects of investigations were the young cherry trees obtained as a result of budding of mahaleb cherry (Prunus mahaleb L. and sweet cherry (Prunus avium L. seedlings in the way by the chip budding-15th July and T-budding-on the 15th July and the 1st September. The used terms and budding methods did not affect the bud taking and the quality of cherry trees during three years studies. Chip budding of the sweet cherry on the 15th July was the most effective way of this seedling budding. Late budding-on the 1st September-did not change the efficiency of the nursery only in case of mahaleb cherry. The highest number-33 000 of the young trees, average per 1 ha was got as a result of the chip and "T" mahaleb cherry budding on the 1st September.

  19. Change in Auxin and Cytokinin Levels Coincides with Altered Expression of Branching Genes during Axillary Bud Outgrowth in Chrysanthemum.

    Science.gov (United States)

    Dierck, Robrecht; De Keyser, Ellen; De Riek, Jan; Dhooghe, Emmy; Van Huylenbroeck, Johan; Prinsen, Els; Van Der Straeten, Dominique

    2016-01-01

    In the production and breeding of Chrysanthemum sp., shoot branching is an important quality aspect as the outgrowth of axillary buds determines the final plant shape. Bud outgrowth is mainly controlled by apical dominance and the crosstalk between the plant hormones auxin, cytokinin and strigolactone. In this work the hormonal and genetic regulation of axillary bud outgrowth was studied in two differently branching cut flower Chrysanthemum morifolium (Ramat) genotypes. C17 is a split-type which forms an inflorescence meristem after a certain vegetative period, while C18 remains vegetative under long day conditions. Plant growth of both genotypes was monitored during 5 subsequent weeks starting one week before flower initiation occurred in C17. Axillary bud outgrowth was measured weekly and samples of shoot apex, stem and axillary buds were taken during the first two weeks. We combined auxin and cytokinin measurements by UPLC-MS/MS with RT-qPCR expression analysis of genes involved in shoot branching regulation pathways in chrysanthemum. These included bud development genes (CmBRC1, CmDRM1, CmSTM, CmLsL), auxin pathway genes (CmPIN1, CmTIR3, CmTIR1, CmAXR1, CmAXR6, CmAXR2, CmIAA16, CmIAA12), cytokinin pathway genes (CmIPT3, CmHK3, CmRR1) and strigolactone genes (CmMAX1 and CmMAX2). Genotype C17 showed a release from apical dominance after floral transition coinciding with decreased auxin and increased cytokinin levels in the subapical axillary buds. As opposed to C17, C18 maintained strong apical dominance with vegetative growth throughout the experiment. Here high auxin levels and decreasing cytokinin levels in axillary buds and stem were measured. A differential expression of several branching genes accompanied the different hormonal change and bud outgrowth in C17 and C18. This was clear for the strigolactone biosynthesis gene CmMAX1, the transcription factor CmBRC1 and the dormancy associated gene CmDRM1, that all showed a decreased expression in C17 at floral

  20. Change in Auxin and Cytokinin Levels Coincides with Altered Expression of Branching Genes during Axillary Bud Outgrowth in Chrysanthemum.

    Directory of Open Access Journals (Sweden)

    Robrecht Dierck

    Full Text Available In the production and breeding of Chrysanthemum sp., shoot branching is an important quality aspect as the outgrowth of axillary buds determines the final plant shape. Bud outgrowth is mainly controlled by apical dominance and the crosstalk between the plant hormones auxin, cytokinin and strigolactone. In this work the hormonal and genetic regulation of axillary bud outgrowth was studied in two differently branching cut flower Chrysanthemum morifolium (Ramat genotypes. C17 is a split-type which forms an inflorescence meristem after a certain vegetative period, while C18 remains vegetative under long day conditions. Plant growth of both genotypes was monitored during 5 subsequent weeks starting one week before flower initiation occurred in C17. Axillary bud outgrowth was measured weekly and samples of shoot apex, stem and axillary buds were taken during the first two weeks. We combined auxin and cytokinin measurements by UPLC-MS/MS with RT-qPCR expression analysis of genes involved in shoot branching regulation pathways in chrysanthemum. These included bud development genes (CmBRC1, CmDRM1, CmSTM, CmLsL, auxin pathway genes (CmPIN1, CmTIR3, CmTIR1, CmAXR1, CmAXR6, CmAXR2, CmIAA16, CmIAA12, cytokinin pathway genes (CmIPT3, CmHK3, CmRR1 and strigolactone genes (CmMAX1 and CmMAX2. Genotype C17 showed a release from apical dominance after floral transition coinciding with decreased auxin and increased cytokinin levels in the subapical axillary buds. As opposed to C17, C18 maintained strong apical dominance with vegetative growth throughout the experiment. Here high auxin levels and decreasing cytokinin levels in axillary buds and stem were measured. A differential expression of several branching genes accompanied the different hormonal change and bud outgrowth in C17 and C18. This was clear for the strigolactone biosynthesis gene CmMAX1, the transcription factor CmBRC1 and the dormancy associated gene CmDRM1, that all showed a decreased expression in

  1. Transcriptome sequencing and identification of cold tolerance genes in hardy Corylus species (C. heterophylla Fisch) floral buds.

    Science.gov (United States)

    Chen, Xin; Zhang, Jin; Liu, Qingzhong; Guo, Wei; Zhao, Tiantian; Ma, Qinghua; Wang, Guixi

    2014-01-01

    The genus Corylus is an important woody species in Northeast China. Its products, hazelnuts, constitute one of the most important raw materials for the pastry and chocolate industry. However, limited genetic research has focused on Corylus because of the lack of genomic resources. The advent of high-throughput sequencing technologies provides a turning point for Corylus research. In the present study, we performed de novo transcriptome sequencing for the first time to produce a comprehensive database for the Corylus heterophylla Fisch floral buds. The C. heterophylla Fisch floral buds transcriptome was sequenced using the Illumina paired-end sequencing technology. We produced 28,930,890 raw reads and assembled them into 82,684 contigs. A total of 40,941 unigenes were identified, among which 30,549 were annotated in the NCBI Non-redundant (Nr) protein database and 18,581 were annotated in the Swiss-Prot database. Of these annotated unigenes, 25,311 and 10,514 unigenes were assigned to gene ontology (GO) categories and clusters of orthologous groups (COG), respectively. We could map 17,207 unigenes onto 128 pathways using the Kyoto Encyclopedia of Genes and Genomes Pathway (KEGG) database. Additionally, based on the transcriptome, we constructed a candidate cold tolerance gene set of C. heterophylla Fisch floral buds. The expression patterns of selected genes during four stages of cold acclimation suggested that these genes might be involved in different cold responsive stages in C. heterophylla Fisch floral buds. The transcriptome of C. heterophylla Fisch floral buds was deep sequenced, de novo assembled, and annotated, providing abundant data to better understand the C. heterophylla Fisch floral buds transcriptome. Candidate genes potentially involved in cold tolerance were identified, providing a material basis for future molecular mechanism analysis of C. heterophylla Fisch floral buds tolerant to cold stress.

  2. Prognostic and diagnostic significance of tumor budding associated with β-catenin expression in submucosal invasive colorectal carcinoma.

    Science.gov (United States)

    Umemura, Ken; Takagi, Sho; Shimada, Takenobu; Masuda, Takayuki; Shiga, Hisashi; Takahashi, Shuichiro; Takahashi, Seiichi; Kinouchi, Yoshitaka; Shibuya, Daisuke; Shimosegawa, Tooru

    2013-01-01

    Endoscopic resection has become a major curative treatment for early colorectal carcinoma without lymph node metastasis. However, lymph node metastasis, a poor prognostic factor in colorectal carcinoma, occurs in about 10% of the patients with submucosal invasive colorectal carcinoma. Therefore, it is important to identify a high-risk factor for lymph node metastasis in submucosal invasive colorectal carcinoma. This study was designed to identify the relationship between tumor budding with β-catenin expression and lymph node metastasis in submucosal invasive colorectal carcinoma. We investigated the immunohistochemistry of tumor budding in the 142 patients who underwent surgical resection for submucosal invasive colorectal carcinomas between 1984 and 1999 and the expression pattern of β-catenin in budding tumor cells. Accordingly, all the patients were followed up for at least 10 years or until death. Among the 142 patients, lymph node metastasis was detected in 14 patients (9.9%). Univariate analysis showed that tumor budding with ≥ 5 tumor cells or cell clusters with expression of β-catenin in the nucleus was significantly associated with lymph node metastasis (P = 0.005). In contrast, tumor budding detected by hematoxylin and eosin staining was not associated with lymph node metastasis. Multivariate logistic regression analysis showed that tumor budding with ≥ 5 tumor cells or cell clusters with expression of β-catenin in the nucleus was a significant risk factor for lymph node metastasis (odds ratio, 7.124; 95% confidence interval, 1.407-36.062). Thus, tumor budding associated with β-catenin expression is a risk factor for lymph node metastasis in submucosal invasive colorectal carcinoma.

  3. The rRNA methyltransferase Bud23 shows functional interaction with components of the SSU processome and RNase MRP.

    Science.gov (United States)

    Sardana, Richa; White, Joshua P; Johnson, Arlen W

    2013-06-01

    Bud23 is responsible for the conserved methylation of G1575 of 18S rRNA, in the P-site of the small subunit of the ribosome. bud23Δ mutants have severely reduced small subunit levels and show a general failure in cleavage at site A2 during rRNA processing. Site A2 is the primary cleavage site for separating the precursors of 18S and 25S rRNAs. Here, we have taken a genetic approach to identify the functional environment of BUD23. We found mutations in UTP2 and UTP14, encoding components of the SSU processome, as spontaneous suppressors of a bud23Δ mutant. The suppressors improved growth and subunit balance and restored cleavage at site A2. In a directed screen of 50 ribosomal trans-acting factors, we identified strong positive and negative genetic interactions with components of the SSU processome and strong negative interactions with components of RNase MRP. RNase MRP is responsible for cleavage at site A3 in pre-rRNA, an alternative cleavage site for separating the precursor rRNAs. The strong negative genetic interaction between RNase MRP mutants and bud23Δ is likely due to the combined defects in cleavage at A2 and A3. Our results suggest that Bud23 plays a role at the time of A2 cleavage, earlier than previously thought. The genetic interaction with the SSU processome suggests that Bud23 could be involved in triggering disassembly of the SSU processome, or of particular subcomplexes of the processome.

  4. Constitutive expression of the Poplar FD-like basic leucine zipper transcription factor alters growth and bud development.

    Science.gov (United States)

    Parmentier-Line, Cécile M; Coleman, Gary D

    2016-01-01

    In poplar, the CO/FT regulatory module mediates seasonal growth cessation. Although FT interacts with the basic leucine zipper transcription factor FD, surprisingly little is known about the possible role of FD in bud development and growth cessation in trees. In this study, we examined the expression and localization of the poplar FD homolog, PtFD1, during short-day (SD)-induced bud development, and the consequences of overexpressing PtFD1 on bud development and shoot growth. PtFD1 was primarily expressed in apical and axillary buds and exhibited a transient increase in expression during the initial stages of SD-induced bud development. This transient increase declined with continued SD treatment. When PtFD1 was overexpressed in poplar, SD-induced growth cessation and bud formation were abolished. PTFD1 overexpression also resulted in precocious flowering of juvenile plants in long-day (LD) photoperiods. Because the phenotypes associated with overexpression of PtFD1 are similar to those observe when poplar FT1 is overexpressed (Science, 312, 2006, 1040), the expression and diurnal patterns of expression of both poplar FT1 and FT2 were characterized in PtFD1 overexpression poplars and found to be altered. DNA microarray analysis revealed few differences in gene expression between PtFD1 overexpressing poplars in LD conditions while extensive levels of differential gene expression occur in SD-treated plants. These results enforce the connection between the regulation of flowering and the regulation of growth cessation and bud development in poplar. © 2015 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

  5. Molecular profiling of tumour budding implicates TGFβ-mediated epithelial–mesenchymal transition as a therapeutic target in oral squamous cell carcinoma

    DEFF Research Database (Denmark)

    Jensen, David Hebbelstrup; Dabelsteen, Erik; Specht, Lena

    2015-01-01

    analysis to demonstrate that the number of tumour buds in cytokeratin-stained sections correlates with patients having lymph node metastases at diagnosis. The tumour bud count was also a predictor of overall survival, independent of TNM stage. Tumour buds and paired central tumour areas were subsequently...... collected from oral squamous cell carcinoma (OSCC) specimens using laser capture microdissection and examined with RNA sequencing and miRNA-qPCR arrays. Compared with cells from the central parts of the tumours, budding cells exhibited a particular gene expression signature comprising factors involved....... Moreover, miR-200 family members were downregulated in budding tumour cells. We used immunohistochemistry to validate five markers of the EMT/MET process in 199 OSCC tumours, as well as in situ hybridization in 20 OSCC samples. Given the strong relationship between tumour budding and the development...

  6. Organogenesis in the budding process of the freshwater bryozoan Cristatella mucedo Cuvier, 1798 (Bryozoa, Phylactolaemata).

    Science.gov (United States)

    Schwaha, Thomas; Handschuh, Stephan; Redl, Emanuel; Walzl, Manfred G

    2011-03-01

    The phylogenetic position of bryozoans has been disputed for decades, and molecular phylogenetic analyzes have not unequivocally clarified their position within the Bilateria. As probably the most basal bryozoans, Phylactolaemata is the most promising taxon for large-scale phylogenetic comparisons. These comparisons require extending the morphological and developmental data by investigating different phylactolaemate species to identify basal characters and resolve in-group phylogeny. Accordingly, we analyzed the bud development and the organogenesis of the freshwater bryozoan Cristatella mucedo, with special focus on the formation of the digestive tract and differentiation of the coelomic compartments. Most parts of the digestive tract are formed as an outpocketing at the future anal side growing towards the mouth area. The ganglion is formed by an invagination between the anlagen of the mouth and anus. The lophophoral arms develop as paired lateral protrusions into the lumen of the bud and are temporarily connected by a median, thin bridge. All coelomic compartments are confluent during their development and also in the adult. The epistome coelom develops by fusion of two peritoneal infolds between the gut loop and overgrows the ganglion medially. The coelomic ring canal on the oral side develops by two lateral ingrowths and supplies the oral tentacles. On the forked canal, supplying the innermost row of tentacles above the epistome, a bladder-shaped swelling, probably with excretory function, is present in some adults. It remains difficult to draw comparisons to other phyla because only few studies have dealt with budding of potentially related taxa in more detail. Nonetheless, our results show that comparative organogenesis can contribute to phylactolaemate systematics and, when more data are available, possibly to that of other bryozoan classes and bilaterian phyla. Copyright © 2010 Wiley-Liss, Inc.

  7. Screening the budding yeast genome reveals unique factors affecting K2 toxin susceptibility.

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    Elena Servienė

    Full Text Available BACKGROUND: Understanding how biotoxins kill cells is of prime importance in biomedicine and the food industry. The budding yeast (S. cerevisiae killers serve as a convenient model to study the activity of biotoxins consistently supplying with significant insights into the basic mechanisms of virus-host cell interactions and toxin entry into eukaryotic target cells. K1 and K2 toxins are active at the cell wall, leading to the disruption of the plasma membrane and subsequent cell death by ion leakage. K28 toxin is active in the cell nucleus, blocking DNA synthesis and cell cycle progression, thereby triggering apoptosis. Genome-wide screens in the budding yeast S. cerevisiae identified several hundred effectors of K1 and K28 toxins. Surprisingly, no such screen had been performed for K2 toxin, the most frequent killer toxin among industrial budding yeasts. PRINCIPAL FINDINGS: We conducted several concurrent genome-wide screens in S. cerevisiae and identified 332 novel K2 toxin effectors. The effectors involved in K2 resistance and hypersensitivity largely map in distinct cellular pathways, including cell wall and plasma membrane structure/biogenesis and mitochondrial function for K2 resistance, and cell wall stress signaling and ion/pH homeostasis for K2 hypersensitivity. 70% of K2 effectors are different from those involved in K1 or K28 susceptibility. SIGNIFICANCE: Our work demonstrates that despite the fact that K1 and K2 toxins share some aspects of their killing strategies, they largely rely on different sets of effectors. Since the vast majority of the host factors identified here is exclusively active towards K2, we conclude that cells have acquired a specific K2 toxin effectors set. Our work thus indicates that K1 and K2 have elaborated different biological pathways and provides a first step towards the detailed characterization of K2 mode of action.

  8. Chronic Oral Capsaicin Exposure During Development Leads to Adult Rats with Reduced Taste Bud Volumes

    Science.gov (United States)

    Omelian, Jacquelyn M.; Samson, Kaeli K.; Sollars, Suzanne I.

    2016-01-01

    Introduction Cross-sensory interaction between gustatory and trigeminal nerves occurs in the anterior tongue. Surgical manipulations have demonstrated that the strength of this relationship varies across development. Capsaicin is a neurotoxin that affects fibers of the somatosensory lingual nerve surrounding taste buds, but not fibers of the gustatory chorda tympani nerve which synapse with taste receptor cells. Since capsaicin is commonly consumed by many species, including humans, experimental use of this neurotoxin provides a naturalistic perturbation of the lingual trigeminal system. Neonatal or adults rats consumed oral capsaicin for 40 days and we examined the cross-sensory effect on the morphology of taste buds across development. Methods Rats received moderate doses of oral capsaicin, with chronic treatments occurring either before or after taste system maturation. Tongue morphology was examined either 2 or 50 days after treatment cessation. Edema, which has been previously suggested as a cause of changes in capsaicin-related gustatory function, was also assessed. Results Reductions in taste bud volume occurred 50 days, but not 2 days post-treatment for rats treated as neonates. Adult rats at either time post-treatment were unaffected. Edema was not found to occur with the 5 ppm concentration of capsaicin we used. Conclusions Results further elucidate the cooperative relationship between these discrete sensory systems and highlight the developmentally mediated aspect of this interaction. Implications Chronic exposure to even moderate levels of noxious stimuli during development has the ability to impact the orosensory environment, and these changes may not be evident until long after exposure has ceased. PMID:28083080

  9. Discovery of an unconventional centromere in budding yeast redefines evolution of point centromeres.

    Science.gov (United States)

    Kobayashi, Norihiko; Suzuki, Yutaka; Schoenfeld, Lori W; Müller, Carolin A; Nieduszynski, Conrad; Wolfe, Kenneth H; Tanaka, Tomoyuki U

    2015-08-03

    Centromeres are the chromosomal regions promoting kinetochore assembly for chromosome segregation. In many eukaryotes, the centromere consists of up to mega base pairs of DNA. On such "regional centromeres," kinetochore assembly is mainly defined by epigenetic regulation [1]. By contrast, a clade of budding yeasts (Saccharomycetaceae) has a "point centromere" of 120-200 base pairs of DNA, on which kinetochore assembly is defined by the consensus DNA sequence [2, 3]. During evolution, budding yeasts acquired point centromeres, which replaced ancestral, regional centromeres [4]. All known point centromeres among different yeast species share common consensus DNA elements (CDEs) [5, 6], implying that they evolved only once and stayed essentially unchanged throughout evolution. Here, we identify a yeast centromere that challenges this view: that of the budding yeast Naumovozyma castellii is the first unconventional point centromere with unique CDEs. The N. castellii centromere CDEs are essential for centromere function but have different DNA sequences from CDEs in other point centromeres. Gene order analyses around N. castellii centromeres indicate their unique, and separate, evolutionary origin. Nevertheless, they are still bound by the ortholog of the CBF3 complex, which recognizes CDEs in other point centromeres. The new type of point centromere originated prior to the divergence between N. castellii and its close relative Naumovozyma dairenensis and disseminated to all N. castellii chromosomes through extensive genome rearrangement. Thus, contrary to the conventional view, point centromeres can undergo rapid evolutionary changes. These findings give new insights into the evolution of point centromeres. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  10. Transcriptome Analysis of Floral Buds Deciphered an Irregular Course of Meiosis in Polyploid Brassica rapa.

    Science.gov (United States)

    Braynen, Janeen; Yang, Yan; Wei, Fang; Cao, Gangqiang; Shi, Gongyao; Tian, Baoming; Zhang, Xiaowei; Jia, Hao; Wei, Xiaochun; Wei, Zhenzhen

    2017-01-01

    Polyploidy is a fundamental process in plant evolution. Understanding the polyploidy-associated effects on plant reproduction is essential for polyploid breeding program. In the present study, our cytological analysis firstly demonstrated that an overall course of meiosis was apparently distorted in the synthetic polyploid Brassica rapa in comparison with its diploid progenitor. To elucidate genetic basis of this irregular meiosis at a molecular level, the comparative RNA-seq analysis was further used to investigate differential genetic regulation of developing floral buds identified at meiosis between autotetraploid and diploid B. rapa. In total, compared to its diploid counterparts, among all 40,927 expressed genes revealed, 4,601 differentially expressed genes (DEGs) were identified in the floral buds of autotetraploid B. rapa, among which 288 DEGs annotated were involved in meiosis. Notably, DMC1 identified as one previously known meiosis-specific gene involved in inter-homologous chromosome dependent repair of DNA double stranded breaks (DSBs), was significantly down-regulated in autotetraploid B. rapa, which presumably contributed to abnormal progression during meiosis I. Although certain DEGs associated with RNA helicase, cell cycling, and somatic DNA repair were up-regulated after genome duplication, genes associated with meiotic DSB repair were significantly down-regulated. Furthermore, the expression of randomly selected DEGs by RNA-seq analysis was confirmed by quantitative real-time PCR analysis in both B. rapa and Arabidopsis thaliana. Our results firstly account for adverse effects of polyploidy on an entire course of meiosis at both cytological and transcriptomic levels, and allow for a comprehensive understanding of the uniformity and differences in the transcriptome of floral buds at meiosis between diploid and polyploid B. rapa as well.

  11. Structure of the archaeal Kae1/Bud32 fusion protein MJ1130: a model for the eukaryotic EKC/KEOPS subcomplex.

    Science.gov (United States)

    Hecker, Arnaud; Lopreiato, Raffaele; Graille, Marc; Collinet, Bruno; Forterre, Patrick; Libri, Domenico; van Tilbeurgh, Herman

    2008-09-03

    The EKC/KEOPS yeast complex is involved in telomere maintenance and transcription. The Bud32p and kinase-associated endopeptidase 1 (Kaelp) components of the complex are totally conserved in eukarya and archaea. Their genes are fused in several archaeal genomes, suggesting that they physically interact. We report here the structure of the Methanocaldococcus jannaschii Kae1/Bud32 fusion protein MJ1130. Kae1 is an iron protein with an ASKHA fold and Bud32 is an atypical small RIO-type kinase. The structure MJ1130 suggests that association with Kae1 maintains the Bud32 kinase in an inactive state. We indeed show that yeast Kae1p represses the kinase activity of yeast Bud32p. Extensive conserved interactions between MjKae1 and MjBud32 suggest that Kae1p and Bud32p directly interact in both yeast and archaea. Mutations that disrupt the Kae1p/Bud32p interaction in the context of the yeast complex have dramatic effects in vivo and in vitro, similar to those observed with deletion mutations of the respective components. Direct interaction between Kae1p and Bud32p in yeast is required both for the transcription and the telomere homeostasis function of EKC/KEOPS.

  12. Regulation of RhSUC2, a sucrose transporter, is correlated with the light control of bud burst in Rosa sp.

    Science.gov (United States)

    Henry, Clemence; Rabot, Amelie; Laloi, Maryse; Mortreau, Eric; Sigogne, Monique; Leduc, Nathalie; Lemoine, Rémi; Sakr, Soulaiman; Vian, Alain; Pelleschi-Travier, Sandrine

    2011-10-01

    In roses, light is a central environmental factor controlling bud break and involves a stimulation of sugar metabolism. Very little is known about the role of sucrose transporters in the bud break process and its regulation by light. In this study, we show that sugar promotes rose bud break and that bud break is accompanied by an import of sucrose. Radio-labelled sucrose accumulation is higher in buds exposed to light than to darkness and involves an active component. Several sucrose transporter (RhSUC1, 2, 3 and 4) transcripts are expressed in rose tissues, but RhSUC2 transcript level is the only one induced in buds exposed to light after removing the apical dominance. RhSUC2 is preferentially expressed in bursting buds and stems. Functional analyses in baker's yeast demonstrate that RhSUC2 encodes a sucrose/proton co-transporter with a K(m) value of 2.99 mm at pH 4.5 and shows typical features of sucrose symporters. We therefore propose that bud break photocontrol partly depends upon the modulation of sucrose import into buds by RhSUC2. © 2011 Blackwell Publishing Ltd.

  13. Epithelial-mesenchymal transition (EMT) protein expression in a cohort of stage II colorectal cancer patients with characterized tumor budding and mismatch repair protein status.

    Science.gov (United States)

    Kevans, David; Wang, Lai Mun; Sheahan, Kieran; Hyland, John; O'Donoghue, Diarmuid; Mulcahy, Hugh; O'Sullivan, Jacintha

    2011-12-01

    The relationship between tumor budding, epithelial-mesenchymal transition (EMT) protein expression, and survival has not been closely examined in stage II colorectal cancer (CRC). This study aimed to assess proteins implicated in EMT and to correlate their expression with tumor budding, microsatellite status, and survival. A total of 258 stage II CRCs were identified (tumor budding characterized in 122 cases). Immunohistochemistry for LAMC2, E cadherin, cathepsin L, and β catenin using tissue microarrays was performed. EMT and mismatch repair (MMR) protein expression were correlated with tumor budding and survival. LAMC2 positivity (P tumor budding. In a univariate survival analysis, tumor budding (P tumor budding to be the only variable independently associated with survival: hazard ratio = 7.9 (95% confidence interval = 3-21); P Tumor budding was more frequent in microsatellite-stable (MSS) versus microsatellite-instable (MSI) tumors: 48% versus 26%, respectively; P = .087. MSS cases exhibited reduced membranous β catenin (P = .002) and increased cytoplasmic and nuclear β catenin (P tumor budding and prognosis in early-stage colorectal cancer and requires further evaluation.

  14. Tumor Budding Correlates With the Protumor Immune Microenvironment and Is an Independent Prognostic Factor for Recurrence of Stage I Lung Adenocarcinoma.

    Science.gov (United States)

    Kadota, Kyuichi; Yeh, Yi-Chen; Villena-Vargas, Jonathan; Cherkassky, Leonid; Drill, Esther N; Sima, Camelia S; Jones, David R; Travis, William D; Adusumilli, Prasad S

    2015-09-01

    Immune cell infiltration associated with tumor capsule disruption and tumor budding has been shown to reflect invasiveness, metastasis, and unfavorable prognosis in colorectal cancer. We investigated the influence of tumor budding on prognosis and its association with the immune microenvironment in lung adenocarcinoma. Tumor slides from resected stage I lung adenocarcinomas were reviewed (n = 524 and n = 514, for training and validation cohorts, respectively) for assessment of tumor budding. CD3+ and forkhead box P3+ (FoxP3+) lymphocytes, CD68+ macrophages, IL-7 receptor, and IL-12 receptor β2 were analyzed using tissue microarrays constructed from tumor and stroma. Probability of recurrence was calculated using the competing risks method. In the training cohort, risk of recurrence for high-grade tumor budding was higher than it was for low-grade tumor budding (32% vs 12%, P Tumor budding stratified the risk of recurrence for acinar-predominant (22% vs 9%, P tumors. Tumor budding was associated with higher stromal FoxP3+ lymphocyte infiltration, higher stromal FoxP3/CD3 risk index, higher tumoral and stromal CD68+ macrophage infiltration, and IL-7 receptor overexpression (P Tumor budding remained independently associated with recurrence on multivariate analysis (hazard ratio, 1.61; P = .008). Tumor budding is an independent prognostic factor of stage I lung adenocarcinoma and correlates with the protumor immune microenvironment. Our findings advocate investigating tumor-immune cell interactions at the invading edge as a biologic driver of tumor aggressiveness.

  15. Immunocytochemical analysis of P2X2 in rat circumvallate taste buds

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    Yang Ruibiao

    2012-05-01

    Full Text Available Abstract Background Our laboratory has shown that classical synapses and synaptic proteins are associated with Type III cells. Yet it is generally accepted that Type II cells transduce bitter, sweet and umami stimuli. No classical synapses, however, have been found associated with Type II cells. Recent studies indicate that the ionotropic purinergic receptors P2X2/P2X3 are present in rodent taste buds. Taste nerve processes express the ionotropic purinergic receptors (P2X2/P2X3. P2X2/P2X3Dbl−/− mice are not responsive to sweet, umami and bitter stimuli, and it has been proposed that ATP acts as a neurotransmitter in taste buds. The goal of the present study is to learn more about the nature of purinergic contacts in rat circumvallate taste buds by examining immunoreactivity to antisera directed against the purinergic receptor P2X2. Results P2X2-like immunoreactivity is present in intragemmal nerve processes in rat circumvallate taste buds. Intense immunoreactivity can also be seen in the subgemmal nerve plexuses located below the basal lamina. The P2X2 immunoreactive nerve processes also display syntaxin-1-LIR. The immunoreactive nerves are in close contact with the IP3R3-LIR Type II cells and syntaxin-1-LIR and/or 5-HT-LIR Type III cells. Taste cell synapses are observed only from Type III taste cells onto P2X2-LIR nerve processes. Unusually large, “atypical” mitochondria in the Type II taste cells are found only at close appositions with P2X2-LIR nerve processes. P2X2 immunogold particles are concentrated at the membranes of nerve processes at close appositions with taste cells. Conclusions Based on our immunofluorescence and immunoelectron microscopical studies we believe that both perigemmal and most all intragemmal nerve processes display P2X2-LIR. Moreover, colloidal gold immunoelectron microscopy indicates that P2X2-LIR in nerve processes is concentrated at sites of close apposition with Type II cells. This supports the hypothesis

  16. Host Cell Plasma Membrane Phosphatidylserine Regulates the Assembly and Budding of Ebola Virus.

    Science.gov (United States)

    Adu-Gyamfi, Emmanuel; Johnson, Kristen A; Fraser, Mark E; Scott, Jordan L; Soni, Smita P; Jones, Keaton R; Digman, Michelle A; Gratton, Enrico; Tessier, Charles R; Stahelin, Robert V

    2015-09-01

    Lipid-enveloped viruses replicate and bud from the host cell where they acquire their lipid coat. Ebola virus, which buds from the plasma membrane of the host cell, causes viral hemorrhagic fever and has a high fatality rate. To date, little has been known about how budding and egress of Ebola virus are mediated at the plasma membrane. We have found that the lipid phosphatidylserine (PS) regulates the assembly of Ebola virus matrix protein VP40. VP40 binds PS-containing membranes with nanomolar affinity, and binding of PS regulates VP40 localization and oligomerization on the plasma membrane inner leaflet. Further, alteration of PS levels in mammalian cells inhibits assembly and egress of VP40. Notably, interactions of VP40 with the plasma membrane induced exposure of PS on the outer leaflet of the plasma membrane at sites of egress, whereas PS is typically found only on the inner leaflet. Taking the data together, we present a model accounting for the role of plasma membrane PS in assembly of Ebola virus-like particles. The lipid-enveloped Ebola virus causes severe infection with a high mortality rate and currently lacks FDA-approved therapeutics or vaccines. Ebola virus harbors just seven genes in its genome, and there is a critical requirement for acquisition of its lipid envelope from the plasma membrane of the human cell that it infects during the replication process. There is, however, a dearth of information available on the required contents of this envelope for egress and subsequent attachment and entry. Here we demonstrate that plasma membrane phosphatidylserine is critical for Ebola virus budding from the host cell plasma membrane. This report, to our knowledge, is the first to highlight the role of lipids in human cell membranes in the Ebola virus replication cycle and draws a clear link between selective binding and transport of a lipid across the membrane of the human cell and use of that lipid for subsequent viral entry. Copyright © 2015, American

  17. Pickling process of capers (Capparis spp. flower buds

    Directory of Open Access Journals (Sweden)

    Özcan, Musa

    1999-04-01

    Full Text Available Middle sized (8 < x < 13 mm buds of Capparis spinosa var. spinosa and C. ovata var. canescens from June in brines containing 5,10,15 and 20% salt and from August in brines of 15% salt, and three different size (x < 8 mm, 8 < x < 13 mm, x > 13 mm buds of C. . ovata var. canescens from June in brines of 15% salt were pickled for two months fermentation. Some chemical and microbiological analyses were done in brines during fermentation. Most suitable salt concentration for lactic acid bacteria (LAB activity were 5% and partly 10%. Acidity, LAB activity, sedimentation and hardness were reduced by increasing bud size in C. ovata. Small buds of C. ovata for pickling product had advantage for colour and flavour, however, more sediment and partly softening showed disadvantage. For both species, pickling time was determined as 40 to 50 days in regard of end-product flavour and odour, brine acidity and pH, and LAB activity.

    Se encurtieron durante dos meses botones florales de tamaño medio (8 < x < 13 mm de Capparis spinosa var. spinosa y C. ovata var. canescens, los recolectados en Junio en salmueras conteniendo 5, 10, 15 y 20% de sal, y los de Agosto en salmueras de 15% de sal; y tres tamaños diferentes (x < 8 mm, 8 < x < 13 mm, X > 13 mm de C. ovata var. canescens de Junio en salmueras de 15% de sal. Se realizaron algunos análisis químicos y microbiológicos durante la fermentación. Las concentraciones de sal más adecuadas para la actividad de las bacterias del ácido láctico (LAB fueron 5% y parcialmente 10%. Acidez, actividad de LAB, sedimentación y firmeza (hardness se redujeron al incrementar el tamaño de las alcaparras de C. ovata. Los tamaños pequeños de C. ovata presentaron en el producto encurtido ventajas en color y sabor, pero desventajas por más sedimento y ablandamiento parcial. El tiempo de encurtido para ambas

  18. Incidence, progression and intensity of Bud Rot in Elaeis guineensis Jacq. in San Lorenzo, Ecuador

    OpenAIRE

    Fernando Rivas Figueroa; Fabricio Moreno; Galán Álvaro Rivera Casignia; Lidcay Herrera Isla; Michel Leiva Mora

    2017-01-01

    BUD rot (BR) is the most serious disease of oil palm in Latin America; in Equator has caused more than 150 million USD of losses. The aim of this work was to determine the incidence, progression and disease intensity of BR in E. guineensis. Incidence and disease progression was determined from data of oil palm enterprises: Palesema, PDA, Palpailón, Energy & Palma y Alespalma during 2006-2013. Disease intensity was determined at 2013. Incidence was 66.75 % and disease intensity was 46 %. Based...

  19. Antiviral activity of leaf-bud gum-resin of Tarenna asiatica

    Directory of Open Access Journals (Sweden)

    Vatsavaya Ramabharathi

    2014-08-01

    Full Text Available The leaf-bud exudate of Tarenna asiatica (Rubiaceae: Ixoroideae, Pavetteae is investigated for its biological activity. The crude benzene extract and corymbosin (pure compound isolated were screened for antiviral activity by using ELISA and PCR methods against animal (blue tongue and chikungunya and plant (papaya ring spot, sesbania mosaic and common bean mosaic viruses. Both corymbosin and benzene extract showed significant antiviral activity though corymbosin was found relatively more potent against the animal and plant viruses tested. This is the first report of antiviral activity for the gum-resin of T. asiatica, so also for the compound corymbosin, against the plant viruses.

  20. A checkpoints capturing timing-robust Boolean model of the budding yeast cell cycle regulatory network

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    Hong Changki

    2012-09-01

    Full Text Available Abstract Background Cell cycle process of budding yeast (Saccharomyces cerevisiae consists of four phases: G1, S, G2 and M. Initiated by stimulation of the G1 phase, cell cycle returns to the G1 stationary phase through a sequence of the S, G2 and M phases. During the cell cycle, a cell verifies whether necessary conditions are satisfied at the end of each phase (i.e., checkpoint since damages of any phase can cause severe cell cycle defect. The cell cycle can proceed to the next phase properly only if checkpoint conditions are met. Over the last decade, there have been several studies to construct Boolean models that capture checkpoint conditions. However, they mostly focused on robustness to network perturbations, and the timing robustness has not been much addressed. Only recently, some studies suggested extension of such models towards timing-robust models, but they have not considered checkpoint conditions. Results To construct a timing-robust Boolean model that preserves checkpoint conditions of the budding yeast cell cycle, we used a model verification technique, ‘model checking’. By utilizing automatic and exhaustive verification of model checking, we found that previous models cannot properly capture essential checkpoint conditions in the presence of timing variations. In particular, such models violate the M phase checkpoint condition so that it allows a division of a budding yeast cell into two before the completion of its full DNA replication and synthesis. In this paper, we present a timing-robust model that preserves all the essential checkpoint conditions properly against timing variations. Our simulation results show that the proposed timing-robust model is more robust even against network perturbations and can better represent the nature of cell cycle than previous models. Conclusions To our knowledge this is the first work that rigorously examined the timing robustness of the cell cycle process of budding yeast with respect

  1. Caffeoyl arbutin and related compounds from the buds of Vaccinium dunalianum.

    Science.gov (United States)

    Zhao, Ping; Tanaka, Takashi; Hirabayashi, Keisuke; Zhang, Ying-Jun; Yang, Chong-Ren; Kouno, Isao

    2008-12-01

    Dunalianosides A-I (1-9), esters of arbutin and related phenolic glucosides, were isolated from the buds of Vaccinium dunalianum Wight (Ericaceae) together with 20 known compounds, and their structures were established on the basis of 1- and 2D NMR spectroscopic evidence. Dunalianosides F-H were dimers of p-hydroxyphenyl 6-O-trans-caffeoyl-beta-D-glucopyranoside (10). The latter was obtained in extraordinary high yield (22% of dry weight), and dunalianoside I (9) was found to be a conjugate of arbutin with an iridoid glucoside.

  2. The step-wise pathway of septin hetero-octamer assembly in budding yeast

    OpenAIRE

    Weems, Andrew; McMurray, Michael

    2017-01-01

    Septin proteins bind guanine nucleotides and form rod-shaped hetero-oligomers. Cells choose from a variety of available septins to assemble distinct hetero-oligomers, but the underlying mechanism was unknown. Using a new in vivo assay, we find that a stepwise assembly pathway produces the two species of budding yeast septin hetero-octamers: Cdc11/Shs1?Cdc12?Cdc3?Cdc10?Cdc10?Cdc3?Cdc12?Cdc11/Shs1. Rapid GTP hydrolysis by monomeric Cdc10 drives assembly of the core Cdc10 homodimer. The extended...

  3. bud suckers

    African Journals Online (AJOL)

    The banana plantain (Musa AAB cv Horn 1) is one of the most important staple crop in the world, particularly in Cote d'ivoire where it plays a major role in the economic system. In traditional plantations. farmers use sizeable suckers, also called bayonets, which, generally, are reduced to a single plant per mother stand at the ...

  4. The influence of sun drying process and prolonged storage on composition of essential oil from clove buds (Syzygium aromaticum)

    Science.gov (United States)

    Hastuti, L. T.; Saepudin, E.; Cahyana, A. H.; Rahayu, D. U. C.; Murni, V. W.; Haib, J.

    2017-07-01

    Clove (Syzygium aromaticum) is native to Indonesia and used as a spice in virtually all of the world's cuisine. Clove bud oil, a yellow liquid, is obtained from distillation of buds. The quality of oil is influenced by origin, post-harvest processing, pre-treatment before distillation, the distillation method, and post-distillation treatment. The objective of this study is to investigate the effect of drying process and prolonged storage on essential oil composition of clove bud from the Tolitoli, Indonesia. To determine the effect of drying, fresh clove bud was dried under sunlight until it reached moisture content 13±1 %. The effect of storage was studied in the oil extracted from clove bud that was stored in laboratory at 25 °C for 4 months. The essential oil of each treatment was obtained by steam distillation and its chemical composition was analyzed by GC/MS. The major components found in fresh and dried clove are as follows: eugenol, eugenyl acetate, and caryophyllene. Percentage of caryophyllene was slightly increase after drying but decrease during storage. While the content of eugenyl acetate decreased during drying and storage, the content of eugenol increased. The drying and storage also affect to the change on minor compounds of essential oil of clove.

  5. Bimolecular Complementation to Visualize Filovirus VP40-Host Complexes in Live Mammalian Cells: Toward the Identification of Budding Inhibitors

    Directory of Open Access Journals (Sweden)

    Yuliang Liu

    2011-01-01

    Full Text Available Virus-host interactions play key roles in promoting efficient egress of many RNA viruses, including Ebola virus (EBOV or “e” and Marburg virus (MARV or “m”. Late- (L- domains conserved in viral matrix proteins recruit specific host proteins, such as Tsg101 and Nedd4, to facilitate the budding process. These interactions serve as attractive targets for the development of broad-spectrum budding inhibitors. A major gap still exists in our understanding of the mechanism of filovirus budding due to the difficulty in detecting virus-host complexes and mapping their trafficking patterns in the natural environment of the cell. To address this gap, we used a bimolecular complementation (BiMC approach to detect, localize, and follow the trafficking patterns of eVP40-Tsg101 complexes in live mammalian cells. In addition, we used the BiMC approach along with a VLP budding assay to test small molecule inhibitors identified by in silico screening for their ability to block eVP40 PTAP-mediated interactions with Tsg101 and subsequent budding of eVP40 VLPs. We demonstrated the potential broad spectrum activity of a lead candidate inhibitor by demonstrating its ability to block PTAP-dependent binding of HIV-1 Gag to Tsg101 and subsequent egress of HIV-1 Gag VLPs.

  6. Chemical profiling of clove bud oil (Syzygium aromaticum) from Toli-Toli and Bali by GC-MS analysis

    Science.gov (United States)

    Sulistyoningrum, A. S.; Saepudin, E.; Cahyana, A. H.; Rahayu, D. U. C.; Amelia, B.; Haib, J.

    2017-07-01

    Indonesia is the largest clove producer in the world. In 2012, total world clove production is 113,215 tons where nearly 71 % (79,250 tons) comes from Indonesia. Although Indonesia is a major producer of clove in the world, research and publications about cloves in this country are scarce and hence knowledge about characteristics of difference varieties of cloves is very limited. The present study was aimed to compare major and minor constituents in clove oil responsible for their flavor based on origin which are cloves from Toli-Toli and Bali. The clove bud oil was isolated from clove bud (Syzygium aromaticum) using steam distillation. The compounds of clove bud oil was analyzed using GC-MS. The major compounds of clove oil were eugenol, caryophyllene, α-humulene and eugenyl acetate with composition 66.37 %, 15.38 %, 1.97 % and 12.99 %, respectively (Toli-Toli) and clove from Bali were 72.34 %, 12.51 %, 2.34 % and 5.33 %, respectively. The unique minor compounds of clove oil from Toli-Toli were (+)-δ-cadinene (0.13 %) and β-caryophylladienol (0.19 %) while in clove oil from Bali were valencene (0.17 %), δ-selinene (0.22 %) and alloaromadendrene (0.24 %). A total of 36 compounds were identified from the clove bud oil Toli-Toli and 38 compounds from the clove bud oil Bali.

  7. Bud structure, position and fate generate various branching patterns along shoots of closely related Rosaceae species: a review

    Science.gov (United States)

    Costes, Evelyne; Crespel, Laurent; Denoyes, Béatrice; Morel, Philippe; Demene, Marie-Noëlle; Lauri, Pierre-Eric; Wenden, Bénédicte

    2014-01-01

    Branching in temperate plants is closely linked to bud fates, either floral or vegetative. Here, we review how the fate of meristematic tissues contained in buds and their position along a shoot imprint specific branching patterns which differ among species. Through examples chosen in closely related species in different genera of the Rosaceae family, a panorama of patterns is apparent. Patterns depend on whether vegetative and floral buds are borne individually or together in mixed buds, develop as the shoot grows or after a rest period, and are located in axillary or terminal positions along the parent shoot. The resulting branching patterns are conserved among varieties in a given species but progressively change with the parent shoot length during plant ontogeny. They can also be modulated by agronomic and environmental conditions. The existence of various organizations in the topology and fate of meristematic tissues and their appendages in closely related species questions the between-species conservation of physiological and molecular mechanisms leading to bud outgrowth vs. quiescence and to floral induction vs. vegetative development. PMID:25520729

  8. A stochastic model correctly predicts changes in budding yeast cell cycle dynamics upon periodic expression of CLN2.

    Directory of Open Access Journals (Sweden)

    Cihan Oguz

    Full Text Available In this study, we focus on a recent stochastic budding yeast cell cycle model. First, we estimate the model parameters using extensive data sets: phenotypes of 110 genetic strains, single cell statistics of wild type and cln3 strains. Optimization of stochastic model parameters is achieved by an automated algorithm we recently used for a deterministic cell cycle model. Next, in order to test the predictive ability of the stochastic model, we focus on a recent experimental study in which forced periodic expression of CLN2 cyclin (driven by MET3 promoter in cln3 background has been used to synchronize budding yeast cell colonies. We demonstrate that the model correctly predicts the experimentally observed synchronization levels and cell cycle statistics of mother and daughter cells under various experimental conditions (numerical data that is not enforced in parameter optimization, in addition to correctly predicting the qualitative changes in size control due to forced CLN2 expression. Our model also generates a novel prediction: under frequent CLN2 expression pulses, G1 phase duration is bimodal among small-born cells. These cells originate from daughters with extended budded periods due to size control during the budded period. This novel prediction and the experimental trends captured by the model illustrate the interplay between cell cycle dynamics, synchronization of cell colonies, and size control in budding yeast.

  9. Isolation and measurement of quercetin glucosides in flower buds of Japanese butterbur (Petasites japonicus subsp. gigantea Kitam.).

    Science.gov (United States)

    Matsuura, Hideyuki; Amano, Midori; Kawabata, Jun; Mizutani, Junya

    2002-07-01

    Three quercetin glucosides were isolated from flower buds of Japanese butterbur (Petasites japonicus subsp. gigantea Kitam.) together with caffeic acid as the ingredients that had DPPH radical scavenging activity, using the DPPH-HPLC method for measuring the radical scavenging activity. These quercetin glucosides were identified as quercetin 3-O-beta-D-glucoside, quercetin 3-O-beta-D-6''-O-acetylglucoside, and rutin, and the amounts of the glucosides in flower buds were also examined by HPLC. The flower buds were harvested from four different sites, the total amount of quercetin glucosides in each site was 100-170 mg/100 g fr. wt., and there were no great differences of the amounts between growing fields.

  10. Chilling-dependent release of seed and bud dormancy in peach associates to common changes in gene expression.

    Directory of Open Access Journals (Sweden)

    Carmen Leida

    Full Text Available Reproductive meristems and embryos display dormancy mechanisms in specialized structures named respectively buds and seeds that arrest the growth of perennial plants until environmental conditions are optimal for survival. Dormancy shows common physiological features in buds and seeds. A genotype-specific period of chilling is usually required to release dormancy by molecular mechanisms that are still poorly understood. In order to find common transcriptional pathways associated to dormancy release, we analyzed the chilling-dependent expression in embryos of certain genes that were previously found related to dormancy in flower buds of peach. We propose the presence of short and long-term dormancy events affecting respectively the germination rate and seedling development by independent mechanisms. Short periods of chilling seem to improve germination in an abscisic acid-dependent manner, whereas the positive effect of longer cold treatments on physiological dwarfing coincides with the accumulation of phenylpropanoids in the seed.

  11. Chilling-Dependent Release of Seed and Bud Dormancy in Peach Associates to Common Changes in Gene Expression

    Science.gov (United States)

    Arbona, Vicent; Gómez-Cadenas, Aurelio; Llácer, Gerardo; Badenes, María Luisa; Ríos, Gabino

    2012-01-01

    Reproductive meristems and embryos display dormancy mechanisms in specialized structures named respectively buds and seeds that arrest the growth of perennial plants until environmental conditions are optimal for survival. Dormancy shows common physiological features in buds and seeds. A genotype-specific period of chilling is usually required to release dormancy by molecular mechanisms that are still poorly understood. In order to find common transcriptional pathways associated to dormancy release, we analyzed the chilling-dependent expression in embryos of certain genes that were previously found related to dormancy in flower buds of peach. We propose the presence of short and long-term dormancy events affecting respectively the germination rate and seedling development by independent mechanisms. Short periods of chilling seem to improve germination in an abscisic acid-dependent manner, whereas the positive effect of longer cold treatments on physiological dwarfing coincides with the accumulation of phenylpropanoids in the seed. PMID:22590512

  12. Paramyxovirus Glycoprotein Incorporation, Assembly and Budding: A Three Way Dance for Infectious Particle Production

    Directory of Open Access Journals (Sweden)

    Farah El Najjar

    2014-08-01

    Full Text Available Paramyxoviruses are a family of negative sense RNA viruses whose members cause serious diseases in humans, such as measles virus, mumps virus and respiratory syncytial virus; and in animals, such as Newcastle disease virus and rinderpest virus. Paramyxovirus particles form by assembly of the viral matrix protein, the ribonucleoprotein complex and the surface glycoproteins at the plasma membrane of infected cells and subsequent viral budding. Two major glycoproteins expressed on the viral envelope, the attachment protein and the fusion protein, promote attachment of the virus to host cells and subsequent virus-cell membrane fusion. Incorporation of the surface glycoproteins into infectious progeny particles requires coordinated interplay between the three viral structural components, driven primarily by the matrix protein. In this review, we discuss recent progress in understanding the contributions of the matrix protein and glycoproteins in driving paramyxovirus assembly and budding while focusing on the viral protein interactions underlying this process and the intracellular trafficking pathways for targeting viral components to assembly sites. Differences in the mechanisms of particle production among the different family members will be highlighted throughout.

  13. Evaluation of extracts and isolated fraction from Capparis spinosa L. buds as an antioxidant source.

    Science.gov (United States)

    Germanò, Maria Paola; De Pasquale, Rita; D'Angelo, Valeria; Catania, Stefania; Silvari, Virginia; Costa, Chiara

    2002-02-27

    The antioxidant activity of extracts from Capparis spinosa L. buds was evaluated using different in vitro tests: ascorbate/Fe(2+)-mediated lipid peroxidation of microsomes from rat liver; bleaching of 1,1-diphenyl-2-picryl-hydrazyl radical; and autoxidation of Fe(2+) ion in the presence of bathophenanthroline disulfonate. The methanolic extract showed strong activities in all of these in vitro tests. The amount of total phenols was determined in the methanolic extract. In addition, the level of rutin was calculated as 0.39% (w/w) by HPLC analysis. Our findings indicate the following: (a) the antioxidant efficiency of the methanolic extract may be attributed to its phenolic content; and (b) the antioxidant activity of the methanolic extract was maintained after removal of glucosinolates, confirming that these compounds do not interfere with the antioxidant properties of the extract. The results obtained from this study exalt the nutritional value of the flowering buds (capers) which are widely used as a source of flavor.

  14. Behavior of lateral buds of Hancornia speciosa after cryopreservation by encapsulation-vitrification

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    Débora de Oliveira Prudente

    2017-05-01

    Full Text Available Hancornia speciosa is a fruitful species from Cerrado biome with high economic potential. However, the intense and disordered extractivism have caused a reduction of its population in its endemic area. In addition, seed recalcitrance negatively affects the conventional conservation of the species. Aiming to find alternatives that enable the long-term conservation of this species, the study’s objective was to assess the behavior of lateral bud’s regrowth after cryopreservation procedures by encapsulation-vitrification technique. Sodium alginate capsules containing lateral buds were pre-cultured in liquid WPM supplemented with 1.0 M glycerol, and subsequently exposed to different concentrations of sucrose (0.3; 0.75 and 1.0 M for 24 or 48 hours. The capsules were subjected to dehydration in silica gel or airflow hood for 0, 1, 2 and 3 hours before different incubation times in PVS2 (0, 15, 30, 60 and 120 minutes at 0°C. A high regeneration percentage of lateral buds was observed after cryopreservation of capsules treated with 0.75 M sucrose plus 1.0 M glycerol (24 hours, associated with dehydration in an airflow hood (1 hour and immersion in PVS2 (15 minutes. Encapsulation-vitrification allowed the long-term conservation, and provided high plant material survival rates after cryopreservation of Hancornia speciosa sensitive explants.

  15. A kinetic study of bitter taste receptor sensing using immobilized porcine taste bud tissues.

    Science.gov (United States)

    Wei, Lihui; Qiao, Lixin; Pang, Guangchang; Xie, Junbo

    2017-06-15

    At present, developing an efficient assay method for truly reflecting the real feelings of gustatory tissues is of great importance. In this study, a novel biosensor was fabricated to investigate the kinetic characteristics of the receptors in taste bud tissues sensing bitter substances for the first time. Porcine taste bud tissues were used as the sensing elements, and the sandwich-type sensing membrane was fixed onto a glassy carbon electrode for assembling the biosensor. With the developed sensor, the response currents induced by sucrose octaacetate, denatonium benzoate, and quercetin stimulating corresponding receptors were determined. The results demonstrated that the interaction between the analyst with their receptors were fitting to hyperbola (R 2 =0.9776, 0.9980 and 0.9601), and the activation constants were 8.748×10 -15 mol/L, 1.429×10 -12 mol/L, 6.613×10 -14 mol/L, respectively. The average number of receptors per cell was calculated as 1.75, 28.58, and 13.23, while the signal amplification factors were 1.08×10 4 , 2.89×10 3 and 9.76×10 4 . These suggest that the sensor can be used to quantitatively describe the interaction characteristics of cells or tissue receptors with their ligands, the role of cellular signaling cascade, the number of receptors, and the signal transmission pathways. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Micropropagation of paradise tree (Melia azedarach) by in vitro culture of axillary buds.

    Science.gov (United States)

    Mroginski, Luis A; Rey, Hebe Y

    2013-01-01

    Paradise tree (Melia azedarach L.) is a multipurpose ornamental and timber tree, and its extracts are used to make insecticides and fungicides. Conventional propagation is done by seeds; however, sexual reproduction results in wide genetic variability. Therefore, clonal propagation is desirable to reduce genetic variation. This chapter describes a protocol for in vitro propagation of paradise tree by axillary buds. There are major steps for this protocol. Firstly, shoot induction by in vitro culture of axillary buds, excised from potted plants obtained by rooting of cuttings of 10-15-year-old adult trees. The initiation medium was composed of Murashige and Skoog medium (MS) supplemented with 0.5 mg/L BAP (benzylaminopurine), 0.1 mg/L IBA (indolebutyric acid), and 0.1 mg/L GA(3) (gibberellic acid). Secondly, multiplication of the regenerated shoots on MS medium amended with 0.5 mg/L BAP and 0.1 mg/L GA(3). Thirdly, rooting of the regenerated shoots on MS medium containing 0.1 mg/L IBA. Fully well-developed plants were transferred to pots containing sand, peat moss, and perlite (1:1:1), and maintained initially in the greenhouse or plastic tunnels.

  17. Negative feedback regulation of auxin signaling by ATHB8/ACL5-BUD2 transcription module.

    Science.gov (United States)

    Baima, Simona; Forte, Valentina; Possenti, Marco; Peñalosa, Andrés; Leoni, Guido; Salvi, Sergio; Felici, Barbara; Ruberti, Ida; Morelli, Giorgio

    2014-06-01

    The role of auxin as main regulator of vascular differentiation is well established, and a direct correlation between the rate of xylem differentiation and the amount of auxin reaching the (pro)cambial cells has been proposed. It has been suggested that thermospermine produced by ACAULIS5 (ACL5) and bushy and dwarf2 (BUD2) is one of the factors downstream to auxin contributing to the regulation of this process in Arabidopsis. Here, we provide an in-depth characterization of the mechanism through which ACL5 modulates xylem differentiation. We show that an increased level of ACL5 slows down xylem differentiation by negatively affecting the expression of homeodomain-leucine zipper (HD-ZIP) III and key auxin signaling genes. This mechanism involves the positive regulation of thermospermine biosynthesis by the HD-ZIP III protein Arabidopsis thaliana homeobox8 tightly controlling the expression of ACL5 and BUD2. In addition, we show that the HD-ZIP III protein REVOLUTA contributes to the increased leaf vascularization and long hypocotyl phenotype of acl5 likely by a direct regulation of auxin signaling genes such as like auxin resistant2 (LAX2) and LAX3. We propose that proper formation and differentiation of xylem depend on a balance between positive and negative feedback loops operating through HD-ZIP III genes. © The Author 2014. Published by the Molecular Plant Shanghai Editorial Office in association with Oxford University Press on behalf of CSPB and IPPE, SIBS, CAS.

  18. Safety assessment of a standardized polyphenolic extract of clove buds: Subchronic toxicity and mutagenicity studies

    Directory of Open Access Journals (Sweden)

    Liju Vijayasteltar

    2016-01-01

    Full Text Available Despite the various reports on the toxicity of clove oil and its major component eugenol, systematic evaluations on the safety of polyphenolic extracts of clove buds have not been reported. Considering the health beneficial pharmacological effects and recent use of clove polyphenols as dietary supplements, the present study investigated the safety of a standardized polyphenolic extract of clove buds (Clovinol, as assessed by oral acute (5 g/kg b.wt. for 14 days and subchronic (0.25, 0.5 and 1 g/kg b.wt. for 90 days toxicity studies on Wistar rats and mutagenicity studies employing Salmonella typhimurium strains. Administration of Clovinol did not result in any toxicologically significant changes in clinical/behavioural observations, ophthalmic examinations, body weights, organ weights, feed consumption, urinalysis, hematology and clinical biochemistry parameters when compared to the untreated control group of animals, indicating the no observed-adverse-effect level (NOAEL as 1000 mg/kg b.wt./day; the highest dose tested. Terminal necropsy did not reveal any treatment-related histopathology changes. Clovinol did not show genotoxicity when tested on TA-98, TA-100 and TA-102 with or without metabolic activation; rather exhibited significant antimutagenic potential against the known mutagens, sodium azide, NPD and tobacco as well as against 2-acetamidoflourene, which needed metabolic activation for mutagenicity.

  19. Presynaptic (Type III) cells in mouse taste buds sense sour (acid) taste.

    Science.gov (United States)

    Huang, Yijen A; Maruyama, Yutaka; Stimac, Robert; Roper, Stephen D

    2008-06-15

    Taste buds contain two types of cells that directly participate in taste transduction - receptor (Type II) cells and presynaptic (Type III) cells. Receptor cells respond to sweet, bitter and umami taste stimulation but until recently the identity of cells that respond directly to sour (acid) tastants has only been inferred from recordings in situ, from behavioural studies, and from immunostaining for putative sour transduction molecules. Using calcium imaging on single isolated taste cells and with biosensor cells to identify neurotransmitter release, we show that presynaptic (Type III) cells specifically respond to acid taste stimulation and release serotonin. By recording responses in cells isolated from taste buds and in taste cells in lingual slices to acetic acid titrated to different acid levels (pH), we also show that the active stimulus for acid taste is the membrane-permeant, uncharged acetic acid moiety (CH(3)COOH), not free protons (H(+)). That observation is consistent with the proximate stimulus for acid taste being intracellular acidification, not extracellular protons per se. These findings may also have implications for other sensory receptors that respond to acids, such as nociceptors.

  20. Somatic embryogenesis from bud and leaf explants of date palm (Phoenix dactylifera L.) cv. Najda.

    Science.gov (United States)

    Mazri, Mouaad Amine; Belkoura, Ilham; Meziani, Reda; Mokhless, Boutaïna; Nour, Souad

    2017-05-01

    An efficient regeneration system through somatic embryogenesis was developed for date palm cv. Najda. Adventitious bud and proximal leaf segments cultured on Murashige and Skoog (MS) medium supplemented with various combinations of auxins and cytokinins induced embryogenesis after at least 6 months of culture. Somatic embryogenesis induction seemed correlated with the type of the explant, the induction period and the auxin used. The highest rate of somatic embryogenesis (86.0%) was obtained on bud explants cultured on MS medium supplemented with 45.0 µM 2,4-dichlorophenoxyacetic acid (2,4-D), and 4.5 µM kinetin or 4.5 µM 6-(dimethylallylamino) purine (2iP). Whereas, low levels of embryogenesis were obtained on media supplemented with 1-naphthalene acetic acid (NAA) or 2-naphthoxyacetic acid (NOA). Proximal leaf segments showed somatic embryogenesis only when cultured on media supplemented with 2,4-D or picloram. Statistical analysis revealed significant effects of explant type and plant growth regulators (PGRs) combination on somatic embryogenesis. Somatic embryos were germinated successfully on PGR-free MS medium with or without activated charcoal (50.0-60.0 and 26.6-36.6%, respectively), and 80.0% of plantlets survived after transferring to a glasshouse for 6 months. Our results will be useful for large-scale propagation of date palm cv. Najda, characterized by high fruit quality and bayoud disease resistance.

  1. RNA-seq data from different developmental stages of Rafflesia cantleyi floral buds.

    Science.gov (United States)

    Amini, Safoora; Alias, Halimah; Aizat-Juhari, Mohd Afiq; Mat-Isa, Mohd-Noor; Adam, Jumaat Haji; Goh, Hoe-Han; Wan, Kiew-Lian

    2017-12-01

    Rafflesia cantleyi, known as one of the world's largest flowers, is a specialised holoparasite due to dramatic morphological modifications. It possesses highly reduced vegetative structure and only appears as a flower for sexual reproduction. Moreover, it has an unusual life cycle in that its floral bud development takes up to nine months. In order to fully understand the highly modified floral organ structure and long life cycle of R. cantleyi, we used Illumina sequencing technology (HiSeq) for sequence generation followed by de novo assembly of sequence reads. We obtained the RNA-seq data from three different stages of floral bud, representing the early, mid and advanced developmental stages. These data are available via BioProject accession number PRJNA378435. More than 10.3 Gb raw sequence data were generated, corresponding to 102,203,042 raw reads. Following removal of low-quality reads and trimming of adapter sequences, a total of 91,638,836 reads were obtained. De novo assembly of these sequences using Trinity resulted in 89,690 unique transcripts with an N50 of 1653 bp. The obtained transcriptomic data will be useful for further study to understand the molecular interactions that result in R. cantleyi floral development.

  2. Cyanogenic Glucosides and Derivatives in Almond and Sweet Cherry Flower Buds from Dormancy to Flowering

    Science.gov (United States)

    Del Cueto, Jorge; Ionescu, Irina A.; Pičmanová, Martina; Gericke, Oliver; Motawia, Mohammed S.; Olsen, Carl E.; Campoy, José A.; Dicenta, Federico; Møller, Birger L.; Sánchez-Pérez, Raquel

    2017-01-01

    Almond and sweet cherry are two economically important species of the Prunus genus. They both produce the cyanogenic glucosides prunasin and amygdalin. As part of a two-component defense system, prunasin and amygdalin release toxic hydrogen cyanide upon cell disruption. In this study, we investigated the potential role within prunasin and amygdalin and some of its derivatives in endodormancy release of these two Prunus species. The content of prunasin and of endogenous prunasin turnover products in the course of flower development was examined in five almond cultivars – differing from very early to extra-late in flowering time – and in one sweet early cherry cultivar. In all cultivars, prunasin began to accumulate in the flower buds shortly after dormancy release and the levels dropped again just before flowering time. In almond and sweet cherry, the turnover of prunasin coincided with increased levels of prunasin amide whereas prunasin anitrile pentoside and β-D-glucose-1-benzoate were abundant in almond and cherry flower buds at certain developmental stages. These findings indicate a role for the turnover of cyanogenic glucosides in controlling flower development in Prunus species. PMID:28579996

  3. Cyanogenic Glucosides and Derivatives in Almond and Sweet Cherry Flower Buds from Dormancy to Flowering

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    Jorge Del Cueto

    2017-05-01

    Full Text Available Almond and sweet cherry are two economically important species of the Prunus genus. They both produce the cyanogenic glucosides prunasin and amygdalin. As part of a two-component defense system, prunasin and amygdalin release toxic hydrogen cyanide upon cell disruption. In this study, we investigated the potential role within prunasin and amygdalin and some of its derivatives in endodormancy release of these two Prunus species. The content of prunasin and of endogenous prunasin turnover products in the course of flower development was examined in five almond cultivars – differing from very early to extra-late in flowering time – and in one sweet early cherry cultivar. In all cultivars, prunasin began to accumulate in the flower buds shortly after dormancy release and the levels dropped again just before flowering time. In almond and sweet cherry, the turnover of prunasin coincided with increased levels of prunasin amide whereas prunasin anitrile pentoside and β-D-glucose-1-benzoate were abundant in almond and cherry flower buds at certain developmental stages. These findings indicate a role for the turnover of cyanogenic glucosides in controlling flower development in Prunus species.

  4. Astral microtubule pivoting promotes their search for cortical anchor sites during mitosis in budding yeast.

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    Stephan Baumgärtner

    Full Text Available Positioning of the mitotic spindle is crucial for proper cell division. In the budding yeast Saccharomyces cerevisiae, two mechanisms contribute to spindle positioning. In the Kar9 pathway, astral microtubules emanating from the daughter-bound spindle pole body interact via the linker protein Kar9 with the myosin Myo2, which moves the microtubule along the actin cables towards the neck. In the dynein pathway, astral microtubules off-load dynein onto the cortical anchor protein Num1, which is followed by dynein pulling on the spindle. Yet, the mechanism by which microtubules target cortical anchor sites is unknown. Here we quantify the pivoting motion of astral microtubules around the spindle pole bodies, which occurs during spindle translocation towards the neck and through the neck. We show that this pivoting is largely driven by the Kar9 pathway. The microtubules emanating from the daughter-bound spindle pole body pivot faster than those at the mother-bound spindle pole body. The Kar9 pathway reduces the time needed for an astral microtubule inside the daughter cell to start pulling on the spindle. Thus, we propose a new role for microtubule pivoting: By pivoting around the spindle pole body, microtubules explore the space laterally, which helps them search for cortical anchor sites in the context of spindle positioning in budding yeast.

  5. New Flavonol Glucuronides from the Flower Buds of Syzygium aromaticum (Clove).

    Science.gov (United States)

    Ryu, Byeol; Kim, Hye Mi; Lee, Jin Su; Lee, Chan Kyu; Sezirahiga, Jurdas; Woo, Jeong-Hwa; Choi, Jung-Hye; Jang, Dae Sik

    2016-04-20

    Repeated chromatography of the EtOAc-soluble fraction from the 70% EtOH extract of the flower buds of Syzygium aromaticum (clove) led to the isolation and characterization of four new flavonol glucuronides, rhamnetin-3-O-β-d-glucuronide (1), rhamnazin-3-O-β-d-glucuronide (2), rhamnazin-3-O-β-d-glucuronide-6″-methyl ester (3), and rhamnocitrin-3-O-β-d-glucuronide-6″-methyl ester (4), together with 15 flavonoids (5-19) having previously known chemical structures. The structures of the new compounds 1-4 were determined by interpretation of spectroscopic data, particularly by 1D- and 2D-NMR studies. Six flavonoids (6, 7, 9, 14, 18, and 19) were isolated from the flower buds of S. aromaticum for the first time in this study. The flavonoids were examined for their cytotoxicity against human ovarian cancer cells (A2780) using MTT assays. Among the isolates, pachypodol (19) showed the most potent cytotoxicity on A2780 cells with an IC50 value of 8.02 μM.

  6. A new acetophenone glycoside from the flower buds of Syzygium aromaticum (cloves).

    Science.gov (United States)

    Ryu, Byeol; Kim, Hye Mi; Woo, Jeong-Hwa; Choi, Jung-Hye; Jang, Dae Sik

    2016-12-01

    A new acetophenone, 2,4,6-trihydroxy-3-methylacetophenone-2-O-β-d-glucoside (1), together with 21 known compounds; one acetophenone (2), four chromone glycosides (3-6), six phenylpropanoids (7-12), six sesquiterpenoids (13-18), two triterpenoids (19 and 20), one sterol (21), and one tannin (22) were isolated from the flower buds of Syzygium aromaticum (cloves). The structure of the new compound 1 was determined by spectroscopic analyses including 1D-, 2D-NMR and HRMS interpretation. Among the isolates, one acetophenone (2), three phenylpropanoids (10-12), and one sesquiterpenoid (13) were isolated from the flower buds of S. aromaticum for the first time in this study. All the isolates (1-22) were evaluated for their cytotoxicity against human ovarian cancer cells (A2780) using MTT assays. Some of the isolates (5, 6, 9, 15, 17, 19, 20, and 21) showed either moderate or weak cytotoxicity on A2780 cells. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. SNAP-, CLIP- and Halo-tag labelling of budding yeast cells.

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    Franziska Stagge

    Full Text Available Fluorescence microscopy of the localization and the spatial and temporal dynamics of specifically labelled proteins is an indispensable tool in cell biology. Besides fluorescent proteins as tags, tag-mediated labelling utilizing self-labelling proteins as the SNAP-, CLIP-, or the Halo-tag are widely used, flexible labelling systems relying on exogenously supplied fluorophores. Unfortunately, labelling of live budding yeast cells proved to be challenging with these approaches because of the limited accessibility of the cell interior to the dyes. In this study we developed a fast and reliable electroporation-based labelling protocol for living budding yeast cells expressing SNAP-, CLIP-, or Halo-tagged fusion proteins. For the Halo-tag, we demonstrate that it is crucial to use the 6'-carboxy isomers and not the 5'-carboxy isomers of important dyes to ensure cell viability. We report on a simple rule for the analysis of ¹H NMR spectra to discriminate between 6'- and 5'-carboxy isomers of fluorescein and rhodamine derivatives. We demonstrate the usability of the labelling protocol by imaging yeast cells with STED super-resolution microscopy and dual colour live cell microscopy. The large number of available fluorophores for these self-labelling proteins and the simplicity of the protocol described here expands the available toolbox for the model organism Saccharomyces cerevisiae.

  8. Safety assessment of a standardized polyphenolic extract of clove buds: Subchronic toxicity and mutagenicity studies.

    Science.gov (United States)

    Vijayasteltar, Liju; Nair, Gopakumar Gopinathan; Maliakel, Balu; Kuttan, Ramadasan; I M, Krishnakumar

    2016-01-01

    Despite the various reports on the toxicity of clove oil and its major component eugenol, systematic evaluations on the safety of polyphenolic extracts of clove buds have not been reported. Considering the health beneficial pharmacological effects and recent use of clove polyphenols as dietary supplements, the present study investigated the safety of a standardized polyphenolic extract of clove buds (Clovinol), as assessed by oral acute (5 g/kg b.wt. for 14 days) and subchronic (0.25, 0.5 and 1 g/kg b.wt. for 90 days) toxicity studies on Wistar rats and mutagenicity studies employing Salmonella typhimurium strains. Administration of Clovinol did not result in any toxicologically significant changes in clinical/behavioural observations, ophthalmic examinations, body weights, organ weights, feed consumption, urinalysis, hematology and clinical biochemistry parameters when compared to the untreated control group of animals, indicating the no observed-adverse-effect level (NOAEL) as 1000 mg/kg b.wt./day; the highest dose tested. Terminal necropsy did not reveal any treatment-related histopathology changes. Clovinol did not show genotoxicity when tested on TA-98, TA-100 and TA-102 with or without metabolic activation; rather exhibited significant antimutagenic potential against the known mutagens, sodium azide, NPD and tobacco as well as against 2-acetamidoflourene, which needed metabolic activation for mutagenicity.

  9. Bone marrow combined with dental bud cells promotes tooth regeneration in miniature pig model.

    Science.gov (United States)

    Kuo, Tzong-Fu; Lin, Hsin-Chi; Yang, Kai-Chiang; Lin, Feng-Huei; Chen, Min-Huey; Wu, Chang-Chin; Chang, Hao-Hueng

    2011-02-01

    Growth factors and morphogens secreted by bone marrow mesenchymal stem cells (BMSCs) of bone marrow fluid may promote tooth regeneration. Accordingly, a tissue engineering approach was utilized to develop an economical strategy for obtaining the growth factors and morphogens from BMSCs. Unerupted second molar tooth buds harvested from miniature pigs were cultured in vitro to obtain dental bud cells (DBCs). Bone marrow fluid, which contains BMSCs, was collected from the porcine mandible before operation. DBCs suspended in bone marrow fluid were seeded into a gelatin/chondoitin-6-sulfate/hyaluronan tri-copolymer scaffold (GCHT scaffold). The DBCs/bone marrow fluid/GCHT scaffold was autografted into the original alveolar sockets of the pigs. Radiographic and histological examinations were applied to identify the structure of regenerated tooth at 40 weeks postimplantation. The present results showed that one pig developed a complete tooth with crown, root, pulp, enamel, dentin, odontoblast, cementum, blood vessel, and periodontal ligament in indiscriminate shape. Three animals had an unerupted tooth that expressed dentin matrix protein-1, vascular endothelial growth factor, and osteopontin; and two other pigs also had dental-like structure with dentin tubules. This study reveals that DBCs adding bone marrow fluid and a suitable scaffold can promote the tooth regeneration in autogenic cell transplantation. © 2010, Copyright the Authors. Artificial Organs © 2010, International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

  10. Distribution and possible function of an adrenomedullin-like peptide in the developing chick limb bud.

    Science.gov (United States)

    Seghatoleslami, M Reza; Martínez, Alfredo; Cuttitta, Frank; Kosher, Robert A

    2002-01-01

    Adrenomedullin (AM) is a multifunctional peptide that exhibits discrete domains of expression during mouse embryogenesis consistent with a role in regulating growth and differentiation during morphogenesis. Here we report that AM immunoreactivity is present at high levels throughout the apical ectodermal ridge (AER) of the chick limb bud as the AER is directing the outgrowth and patterning of underlying limb mesoderm. Immunostaining is particularly strong along the surfaces of the contiguous cells of the AER. AM immunoreactivity attenuates as the AER regresses and is absent from the distal apical ectoderm of stage 20 limbless mutant limb buds which fail to develop an AER. To explore the possible role of AM in AER activity, we examined the effect of exogenous AM and an AM inhibitor on the in vitro morphogenesis of limb mesoderm, cultured in the presence and absence of the AER. Although exogenous AM cannot substitute for the AER in promoting outgrowth of limb mesoderm in vitro, a specific AM antagonist, AM(22-52), impairs the outgrowth and proliferation of limb mesoderm cultured in the presence of the AER. This is consistent with the possibility that inhibition of endogenous AM activity in the AER impairs the ability of the AER to promote limb morphogenesis. Taken together, these studies suggest that an AM-like molecule may function in an autocrine fashion to regulate some aspect of AER activity.

  11. Phenolic composition of "bud extracts" of Ribes nigrum L., Rosa canina L. and Tilia tomentosa M.

    Science.gov (United States)

    Ieri, Francesca; Innocenti, Marzia; Possieri, Lucia; Gallori, Sandra; Mulinacci, Nadia

    2015-11-10

    Liquid preparations such as tinctures and "bud extracts" are typical products widely used in European countries but which have been poorly studied to date. Our research was focused on obtaining a phytochemical characterization of secondary metabolites of black currant, dog rose and silver linden bud extracts and evaluating the changes of selected chemical markers over time. 16 different preparations have been analyzed by the use of HPLC-DAD-MS. Several flavonols were identified in black currant samples including quercetin derivatives (6.7-30.4mg/L). Also hydroxycinnamic acids, mainly p-coumaric acid derivatives (4.1-38.9mg/L) were identified. Flavonols such as glycosides of quercetin and kaempferol, hydroxycinnamic acids, and ellagitannins were detected in dog rose samples, with gallotannins being the main components (up to 1.7g/L). The Tilia tomentosa extracts contained flavonols (quercetin, kaempferol, apigenin derivatives) as principal components with the exception of a single commercial extract with hydroxycinnamic acids as the most abundant metabolites. In this study, without applying accelerated ageing protocols, the stability over time of these liquid preparations was evaluated for up to 10 months and demonstrated negligible variations. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Effects of poplar buds as an alternative to propolis on postharvest diseases control of strawberry fruits.

    Science.gov (United States)

    Yang, Shuzhen; Zhou, Yefeng; Ye, Junli; Fan, Gang; Peng, Litao; Pan, Siyi

    2016-04-01

    Botrytis cinerea and Rhizopus stolonifer, two main postharvest pathogens, cause great loss of strawberry fruits. Here, the effects of poplar buds extracts, a main plant source for Chinese propolis, on disease control were investigated in vitro and in vivo. The HPLC profile of poplar buds ethanol extract (PBEE) was almost identical to that of propolis ethanol extract (PEE), with the active flavonoids identified as pinocembrin, chrysin and galangin. PBEE exhibited similar inhibitory activities on spore germination of both pathogens compared with PEE, and PBEE also strongly inhibited the mycelial growth of the pathogens. In vivo, PBEE could effectively reduce decay of strawberry fruits stored at 13 °C. Although the weight loss was slightly increased, the contents of total soluble solid, titritable acid, vitamin C and total anthocyanins were significantly higher in PBEE treated fruits than those of the control. PBEE had the similar antifungal activity with propolis and had great potential as an alternative to propolis to control strawberry fruits diseases. © 2015 Society of Chemical Industry.

  13. Salvia officinalis L. induces alveolar bud growing in adult female rat mammary glands.

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    Monsefi, Malihezaman; Abedian, Mehrnaz; Azarbahram, Zahra; Ashraf, Mohammad Javad

    2015-01-01

    In traditional medicine Salvia officinalis (sage) has been used as menstrual cycle regulator. In the present study the effects of sage extract on breast tissue were examined. Fourteen female rats were divided into two groups: 1) Distilled water-treated rats (Con) that were gavaged with 1ml distilled water and 2) Saliva officinalis hydroalcoholic extract (SHE)-treated rats that were gavaged with 30mg/kg/body weight of sage extract for 30 days. The estrus cycle changes were monitored by daily examination of vaginal smear. Whole mounts of right pelvic breast were spread on the slide and stained by carmine. The number of alveolar buds (ABs) type 1 and 2 and lobules of mammary gland were scored. Tissue sections of left pelvic mammary gland were prepared and its histomorphometrical changes were measured. Blood samples were taken from dorsal aorta and estradiol and progesterone concentrations were measured using radioimmunoassay. Estrous cycles decreased significantly in SHE-treated animals. The number of alveolar buds and lobules in mammary gland whole mount of SHE-treated group were higher than the Con group. The number and diameter of ducts in histological section of mammary gland in SHE-treated group increased as compared to the Con group. Sage promotes alveologenesis of mammary glands and it can be used as a lactiferous herb.

  14. Apical dominance in saffron and the involvement of the branching enzymes CCD7 and CCD8 in the control of bud sprouting

    Science.gov (United States)

    2014-01-01

    Background In saffron (Crocus sativus), new corms develop at the base of every shoot developed from the maternal corm, a globular underground storage stem. Since the degree of bud sprouts influences the number and size of new corms, and strigolactones (SLs) suppress growth of pre-formed axillary bud, it was considered appropriate to investigate SL involvement in physiology and molecular biology in saffron. We focused on two of the genes within the SL pathway, CCD7 and CCD8, encoding carotenoid cleavage enzymes required for the production of SLs. Results The CsCCD7 and CsCCD8 genes are the first ones isolated and characterized from a non-grass monocotyledonous plant. CsCCD7 and CsCCD8 expression showed some overlapping, although they were not identical. CsCCD8 was highly expressed in quiescent axillary buds and decapitation dramatically reduced its expression levels, suggesting its involvement in the suppression of axillary bud outgrowth. Furthermore, in vitro experiments showed also the involvement of auxin, cytokinin and jasmonic acid on the sprouting of axillary buds from corms in which the apical bud was removed. In addition, CsCCD8 expression, but not CsCCD7, was higher in the newly developed vascular tissue of axillary buds compared to the vascular tissue of the apical bud. Conclusions We showed that production and transport of auxin in saffron corms could act synergistically with SLs to arrest the outgrowth of the axillary buds, similar to the control of above-ground shoot branching. In addition, jasmonic acid seems to play a prominent role in bud dormancy in saffron. While cytokinins from roots promote bud outgrowth. In addition the expression results of CsCCD8 suggest that SLs could positively regulate procambial activity and the development of new vascular tissues connecting leaves with the mother corm. PMID:24947472

  15. GDNF-independent ureteric budding: role of PI3K-independent activation of AKT and FOSB/JUN/AP-1 signaling

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    James B. Tee

    2013-07-01

    A significant fraction of mice deficient in either glial cell-derived neurotrophic factor (GDNF or its co-receptors (Gfrα1, Ret, undergoes ureteric bud (UB outgrowth leading to the formation of a rudimentary kidney. Previous studies using the isolated Wolffian duct (WD culture indicate that activation of fibroblast growth factor (FGF receptor signaling, together with suppression of BMP/Activin signaling, is critical for GDNF-independent WD budding (Maeshima et al., 2007. By expression analysis of embryonic kidney from Ret(−/− mice, we found the upregulation of several FGFs, including FGF7. To examine the intracellular pathways, we then analyzed GDNF-dependent and GDNF-independent budding in the isolated WD culture. In both conditions, Akt activation was found to be important; however, whereas this occurred through PI3-kinase in GDNF-dependent budding, in the case of GDNF-independent budding, Akt activation was apparently via a PI3-kinase independent mechanism. Jnk signaling and the AP-1 transcription factor complex were also implicated in GDNF-independent budding. FosB, a binding partner of c-Jun in the formation of AP-1, was the most highly upregulated gene in the ret knockout kidney (in which budding had still occurred, and we found that its siRNA-mediated knockdown in isolated WDs also blocked GDNF-independent budding. Taken together with the finding that inhibition of Jnk signaling does not block Akt activation/phosphorylation in GDNF-independent budding, the data support necessary roles for both FosB/Jun/AP-1 signaling and PI3-kinase-independent activation of Akt in GDNF-independent budding. A model is proposed for signaling events that involve Akt and JNK working to regulate GDNF-independent WD budding.

  16. The ratio of red light to far red light alters Arabidopsis axillary bud growth and abscisic acid signalling before stem auxin changes.

    Science.gov (United States)

    Holalu, Srinidhi V; Finlayson, Scott A

    2017-02-01

    Arabidopsis thaliana shoot branching is inhibited by a low red light to far red light ratio (R:FR, an indicator of competition), and by loss of phytochrome B function. Prior studies have shown that phytochrome B deficiency suppresses bud growth by elevating systemic auxin signalling, and that increasing the R:FR promotes the growth of buds suppressed by low R:FR by inhibiting bud abscisic acid (ABA) accumulation and signalling. Here, systemic auxin signalling and bud ABA signalling were examined in the context of rapid bud responses to an increased R:FR. Increasing the R:FR promoted the growth of buds inhibited by a low R:FR within 6 h. Relative to a low R:FR, bud ABA accumulation and signalling in plants given a high R:FR showed a sustained decline within 3 h, prior to increased growth. Main stem auxin levels and signalling showed a weak, transient response. Systemic effects and those localised to the bud were further examined by decapitating plants maintained either under a low R:FR or provided with a high R:FR. Increasing the R:FR promoted bud growth before decapitation, but decapitated plants eventually formed longer branches. The data suggest that rapid responses to an increased R:FR may be mediated by changes in bud ABA physiology, although systemic auxin signalling is necessary for sustained bud repression under a low R:FR. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  17. Cell Budding from Normal Appearing Epithelia: A Predictor of Colorectal Cancer Metastasis?

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    Jiang, Bin; Mason, Jeffrey; Jewett, Anahid; Qian, Jun; Ding, Yijiang; Cho, William CS; Zhang, Xichen; Man, Yan-gao

    2013-01-01

    Background: Colorectal carcinogenesis is believed to be a multi-stage process that originates with a localized adenoma, which linearly progresses to an intra-mucosal carcinoma, to an invasive lesion, and finally to metastatic cancer. This progression model is supported by tissue culture and animal model studies, but it is difficult to reconcile with several well-established observations, principally among these are that up to 25% of early stage (Stage I/II), node-negative colorectal cancer (CRC) develop distant metastasis, and that circulating CRC cells are undetectable in peripheral blood samples of up to 50% of patients with confirmed metastasis, but more than 30% of patients with no detectable metastasis exhibit such cells. The mechanism responsible for this diverse behavior is unknown, and there are no effective means to identify patients with pending, or who are at high risk for, developing metastatic CRC. Novel findings: Our previous studies of human breast and prostate cancer have shown that cancer invasion arises from the convergence of a tissue injury, the innate immune response to that injury, and the presence of tumor stem cells within tumor capsules at the site of the injury. Focal degeneration of a capsule due to age or disease attracts lymphocyte infiltration that degrades the degenerating capsules resulting in the formation of a focal disruption in the capsule, which selectively favors proliferating or “budding” of the underlying tumor stem cells. Our recent studies suggest that lymphocyte infiltration also triggers metastasis by disrupting the intercellular junctions and surface adhesion molecules within the proliferating cell buds causing their dissociation. Then, lymphocytes and tumor cells are conjoined through membrane fusion to form tumor-lymphocyte chimeras (TLCs) that allows the tumor stem cell to avail itself of the lymphocyte's natural ability to migrate and breach cell barriers in order to intravasate and to travel to distant organs

  18. Transcriptomic analysis of ‘Suli’ pear (Pyrus pyrifolia white pear group buds during the dormancy by RNA-Seq

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    Liu Guoqin

    2012-12-01

    Full Text Available Abstract Background Bud dormancy is a critical developmental process that allows perennial plants to survive unfavorable environmental conditions. Pear is one of the most important deciduous fruit trees in the world, but the mechanisms regulating bud dormancy in this species are unknown. Because genomic information for pear is currently unavailable, transcriptome and digital gene expression data for this species would be valuable resources to better understand the molecular and biological mechanisms regulating its bud dormancy. Results We performed de novo transcriptome assembly and digital gene expression (DGE profiling analyses of ‘Suli’ pear (Pyrus pyrifolia white pear group using the Illumina RNA-seq system. RNA-Seq generated approximately 100 M high-quality reads that were assembled into 69,393 unigenes (mean length = 853 bp, including 14,531 clusters and 34,194 singletons. A total of 51,448 (74.1% unigenes were annotated using public protein databases with a cut-off E-value above 10-5. We mainly compared gene expression levels at four time-points during bud dormancy. Between Nov. 15 and Dec. 15, Dec. 15 and Jan. 15, and Jan. 15 and Feb. 15, 1,978, 1,024, and 3,468 genes were differentially expressed, respectively. Hierarchical clustering analysis arranged 190 significantly differentially-expressed genes into seven groups. Seven genes were randomly selected to confirm their expression levels using quantitative real-time PCR. Conclusions The new transcriptomes offer comprehensive sequence and DGE profiling data for a dynamic view of transcriptomic variation during bud dormancy in pear. These data provided a basis for future studies of metabolism during bud dormancy in non-model but economically-important perennial species.

  19. Prognostic and predictive factors in gingivo buccal complex squamous cell carcinoma: role of tumor budding and pattern of invasion.

    Science.gov (United States)

    Manjula, B V; Augustine, Suni; Selvam, Sumithra; Mohan, A Mathan

    2015-03-01

    Invasive tumor front (ITF) is the deepest three to six cell layers or detached tumor cell groups at the advancing edge of the tumor. Tumor budding is defined as presence of isolated single cells or small cell clusters scattered in the stroma ahead of the ITF and is characteristic of aggressive cancer. It is recognized as an adverse prognostic factor in several human cancers like colorectal, oesophageal, laryngeal cancers and more recently tongue cancers. However, the prognostic value of tumor budding has not been reported in GBCSCC. The aim of our study was to evaluate the role of pattern of invasion (POI) at the ITF, Tumor budding and other clinicopathological parameters in predicting nodal metastases and prognosis in GBCSCC. 33 patients with primary GBCSCC were prospectively evaluated at a tertiary care referral centre. Tumor budding and type of POI was examined in detail and data documented. Statistical analyses were carried out to assess the correlation of tumor budding, POI, and other clinicopathologic parameters (stage, grade of the tumor, tumor thickness, PNI, LVI) with nodal metastases and predict prognosis. Cox regression was used for both Univariate and multivariate analysis. Significant predictors of nodal metastases on Univariate analysis were male gender (p = 0.021), smoking (p = 0.046), Tumor budding (p = 0.014) and diffuse infiltrative/worst POI (p = 0.004), where as on multivariate analysis only worst POI was significantly associated with positive lymph nodes (p = 0.004). Presence of nodal metastases (p = 0.01) and tumor thickness >5 mm (p = 0.009) were independent negative prognostic factors on multivariate analysis. Significant single risk factor predictive of positive lymph nodes is worst POI in GBCSCC. Nodal metastases and >5 mm tumor thickness are independent risk factors for disease free survival.

  20. Isolation and characterization of the Jatropha curcas APETALA1 (JcAP1) promoter conferring preferential expression in inflorescence buds.

    Science.gov (United States)

    Tao, Yan-Bin; He, Liang-Liang; Niu, Longjian; Xu, Zeng-Fu

    2016-08-01

    The 1.5 kb JcAP1 promoter from the biofuel plant Jatropha curcas is predominantly active in the inflorescence buds of transgenic plants, in which the -1313/-1057 region is essential for maintaining the activity. Arabidopsis thaliana APETALA1 (AP1) is a MADS-domain transcription factor gene that functions primarily in flower development. We isolated a homolog of AP1 from Jatropha curcas (designated JcAP1), which was shown to exhibit flower-specific expression in Jatropha. JcAP1 is first expressed in inflorescence buds and continues to be primarily expressed in the sepals. We isolated a 1.5 kb JcAP1 promoter and evaluated its activity in transgenic Arabidopsis and Jatropha using the β-glucuronidase (GUS) reporter gene. In transgenic Arabidopsis and Jatropha, the inflorescence buds exhibited notable GUS activity, whereas the sepals did not. Against expectations, the JcAP1 promoter was active in the anthers of Arabidopsis and Jatropha and was highly expressed in Jatropha seeds. An analysis of promoter deletions in transgenic Arabidopsis revealed that deletion of the -1313/-1057 region resulted in loss of JcAP1 promoter activity in the inflorescence buds and increased activity in the anthers. These results suggested that some regulatory sequences in the -1313/-1057 region are essential for maintaining promoter activity in inflorescence buds and can partly suppress activity in the anthers. Based on these findings, we hypothesized that other elements located upstream of the 1.5 kb JcAP1 promoter may be required for flower-specific activation. The JcAP1 promoter characterized in this study can be used to drive transgene expression in both the inflorescence buds and seeds of Jatropha.

  1. Tissue culture of horse-radish (Cochlearia armoracia L. meristems: sterilization of buds and comparison of media

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    K. Górecka

    2015-06-01

    Full Text Available The attempt was made to cultivate horse-radish meristems taken from buds removed from roots. The lowest per cent of contamination was found after the buds had been soaiked in 80% ethanol for 6 minutes and then in 5%, 7.5% or 10% chloramine for 30, 30 or 15 minutes, respectively. Both agar media: Murashige-Skoog and Linsmaier-Skoog, were equally good, providing a moderate number of developing plants. The Linsmaier-Skoog medium was more satisfactory when solidified with agar; the results with liquid medium and filter-paper bridges were not as good.

  2. New biofunctional effects of the flower buds of Camellia sinensis and its bioactive acylated oleanane-type triterpene oligoglycosides.

    Science.gov (United States)

    Matsuda, Hisashi; Nakamura, Seikou; Morikawa, Toshio; Muraoka, Osamu; Yoshikawa, Masayuki

    2016-10-01

    We review the biofunctional effects of the flower buds of Camellia sinensis and C. sinensis var. assamica, such as antihyperlipidemic, antihyperglycemic, antiobesity, and gastroprotective effects in vivo, and antiallergic, pancreatic lipase inhibitory, and amyloid β (Aβ) aggregation inhibitory activities in vitro. Although the biofunctional effects of tea leaves have been extensively studied, less attention has been given to those of the flowers and seeds of the tea plant. Our studies focused on the saponin constituents of the extracts of the flower buds of C. sinensis cultivated in Japan and China, and C. sinensis var. assamica cultivated in India, and we review their beneficial biofunctions for health promotion.

  3. High floral bud abscission and lack of open flower abscission in Dendrobium cv. Miss Teen: rapid reduction of ethylene sensitivity in the abscission zone

    NARCIS (Netherlands)

    Bunya-atichart, K.; Ketsa, S.; Doorn, van W.G.

    2006-01-01

    We studied the abscission of floral buds and open flowers in cut Dendrobium inflorescences. Abscission of floral buds was high and sensitive to ethylene in all cultivars studied. Many open flowers abscised in most cultivars, but cv. Willie exhibited only small amount of floral fall and cv. Miss Teen

  4. Unequivocal Identification of 1-Phenylethyl Acetate in Clove Buds (syzygium aromaticum (L. Merr. & L.M.Perry and Clove Essential Oil

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    Klaus Gassenmeier

    2017-06-01

    Full Text Available The natural occurrence of 1-phenylethyl acetate (styrallyl acetate was confirmed in commercially available dried clove buds and also in the hydrodistilled oil from clove buds. This confirms previous reports and other anecdotal evidence for its occurrence in nature.

  5. Tumor budding is an independent risk factor for lymph node metastasis in cutaneous squamous cell carcinoma: a single center retrospective study.

    Science.gov (United States)

    Fujimoto, Masakazu; Yamamoto, Yuki; Matsuzaki, Ibu; Warigaya, Kenji; Iwahashi, Yoshifumi; Kojima, Fumiyoshi; Furukawa, Fukumi; Murata, Shin-Ichi

    2016-09-01

    Although tumor budding is acknowledged as a risk factor for lymph node metastasis in certain types of carcinoma, it is not well investigated in cutaneous squamous cell carcinomas (SCCs). In this study, we analyzed the correlation between tumor budding and nodal metastasis in cutaneous SCC. Histopathologic specimens of 15 metastasizing and 144 non-metastasizing cutaneous SCC were retrospectively reviewed. Cut-off point for tumor budding was determined at five or more foci of an isolated cancer cell or a cluster comprising tumor under hematoxylin and eosin sections. Tumor budding was positive in 93.3% of metastasizing cutaneous SCC and 25.7% of non-metastasizing cutaneous SCC, respectively (p tumor budding was an independent risk factor for nodal metastasis among the other risk factors (tumor size, tumor thickness, Clark level and poor differentiation) (p = 0.0021). In combination with tumor thickness >4 mm, tumor budding became a better marker for predicting nodal metastasis (sensitivity 93.3%, specificity 84.7%). In our cohort, tumor budding was an independent risk factor for nodal metastasis. Our data suggests the promising role of tumor budding in risk evaluation of cutaneous SCC. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  6. Tumor budding and laminin5-γ2 in squamous cell carcinoma of the external auditory canal are associated with shorter survival.

    Science.gov (United States)

    Okado, Yasuko; Aoki, Mikiko; Hamasaki, Makoto; Koga, Kaori; Sueta, Takayuki; Shiratsuchi, Hideki; Oda, Yoshinao; Nakagawa, Takashi; Nabeshima, Kazuki

    2015-01-01

    Squamous cell carcinoma (SCC) of the external auditory canal (EAC) is rare, usually presents at an advanced stage, and is a more aggressive tumor with poor prognosis. The University of Pittsburgh TNM staging system commonly used in prognostication is not perfect, and more accurate biomarkers predicting prognosis are needed. Tumor budding is an established negative prognostic factor at the invasive front in colorectal cancer. Moreover, immunohistochemical studies showed that laminin 5-γ2 (Ln5-γ2) is expressed at the invasive front in tumor or tumor budding cells. We assessed the prognostic significance of tumor budding and Ln5-γ2 expression by performing Ln5-γ2 immunohistochemistry and evaluated the degree of tumor budding in pre-treatment biopsy specimens, and investigated their correlations to clinicopathological parameters in patients with SCC of the EAC. Patients whose tumors had high budding grade and Ln5-γ2 expression had significantly shorter survival times. Budding grade was significantly correlated with Ln5-γ2 expression. Multivariate analysis revealed that high budding grade predicted poorer prognosis regardless of disease stage. Our results suggested that budding grade and Ln5-γ2 expression can be used as indicators of poor prognosis in patients with SCC of the EAC.

  7. Laminin-5 gamma 2 chain expression is associated with intensity of tumor budding and density of stromal myofibroblasts in oral squamous cell carcinoma.

    Science.gov (United States)

    Marangon Junior, Helvécio; Rocha, Valéria Nazaré; Leite, Camila Ferreira; de Aguiar, Maria Cássia Ferreira; Souza, Paulo Eduardo Alencar; Horta, Martinho Campolina Rebello

    2014-03-01

    Oral squamous cell carcinoma (OSCC) is one of the most prevalent cancers worldwide. Laminin-5 gamma 2 chain (laminin-5 γ2) is a protein associated to a migratory phenotype in epithelial neoplastic cells. Stromal myofibroblasts also play a significant role in tumor invasion, due to its ability to modify the extracellular matrix. Tumor budding is a morphologic marker of tumor invasion. The aim of this study was to evaluate the expression of laminin-5 γ2 in OSCC and its association with intensity of tumor budding and density of stromal myofibroblasts. Paraffin-embedded archival samples of 57 OSCC patients were evaluated. Immunohistochemistry was employed to detect laminin-5 γ2, alpha smooth muscle actin (marker of stromal myofibroblasts), and multicytokeratin (to identify OSCC cells in tumor budding evaluation). Laminin-5 γ2 expression and its association with intensity of tumor budding and density of stromal myofibroblasts were analyzed. Association among intensity of tumor budding and density of stromal myofibroblasts was also evaluated. Higher laminin-5 γ2 expression was associated with high-intensity tumor budding (P tumor budding was associated with higher density of stromal myofibroblasts (P tumor budding and with higher density of stromal myofibroblasts, suggesting that this expression is related to the establishment of an invasive phenotype of neoplastic cells and a permissive environment for tumor invasion in this neoplasia. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  8. New report of Lolium multiflorum and Rumex crispus as weed hosts of epiphytic populations of Psuedomonas sp., causal agent of yellow bud in onion in Geogia, USA

    Science.gov (United States)

    Yellow bud, an emerging bacterial disease of onion (Allium cepa L.), has been spreading throughout the Vidalia onion-growing region in Georgia since 2007. Symptoms of yellow bud include intense chlorosis in emerging leaves and severe blight in the older leaves leading to stand loss and reduced bulb ...

  9. Axillary budbreak in a cut rose crop as influenced by light intensity and red:far-red ratio at bud level

    NARCIS (Netherlands)

    Wubs-Timmermans, A.M.; Heuvelink, E.; Marcelis, L.F.M.; Buck-Sorlin, G.H.; Vos, J.

    2014-01-01

    When flower-bearing shoots in cut rose (Rosa ·hybrida) are harvested, a varying number of repressed axillary buds on the shoot remainder start to grow into new shoots (budbreak). Earlier experiments indicated that light reaching the bud affected the number of budbreaks. In all these studies, whole

  10. Expression of ABA Metabolism-Related Genes Suggests Similarities and Differences Between Seed Dormancy and Bud Dormancy of Peach (Prunus persica).

    Science.gov (United States)

    Wang, Dongling; Gao, Zhenzhen; Du, Peiyong; Xiao, Wei; Tan, Qiuping; Chen, Xiude; Li, Ling; Gao, Dongsheng

    2015-01-01

    Dormancy inhibits seed and bud growth of perennial plants until the environmental conditions are optimal for survival. Previous studies indicated that certain co-regulation pathways exist in seed and bud dormancy. In our study, we found that seed and bud dormancy are similar to some extent but show different reactions to chemical treatments that induce breaking of dormancy. Whether the abscisic acid (ABA) regulatory networks are similar in dormant peach seeds and buds is not well known; however, ABA is generally believed to play a critical role in seed and bud dormancy. In peach, some genes putatively involved in ABA synthesis and catabolism were identified and their expression patterns were studied to learn more about ABA homeostasis and the possible crosstalk between bud dormancy and seed dormancy mechanisms. The analysis demonstrated that two 9-cis-epoxycarotenoid dioxygenase-encoding genes seem to be key in regulating ABA biosynthesis to induce seed and bud dormancy. Three CYP707As play an overlapping role in controlling ABA inactivation, resulting in dormancy-release. In addition, Transcript analysis of ABA metabolism-related genes was much similar demonstrated that ABA pathways was similar in the regulation of vegetative and flower bud dormancy, whereas, expression patterns of ABA metabolism-related genes were different in seed dormancy showed that ABA pathway maybe different in regulating seed dormancy in peach.

  11. Ubiquitin conjugation to Gag is essential for ESCRT-mediated HIV-1 budding

    Science.gov (United States)

    2013-01-01

    Background HIV-1 relies on the host ESCRTs for release from cells. HIV-1 Gag engages ESCRTs by directly binding TSG101 or Alix. ESCRTs also sort ubiquitinated membrane proteins through endosomes to facilitate their lysosomal degradation. The ability of ESCRTs to recognize and process ubiquitinated proteins suggests that ESCRT-dependent viral release may also be controlled by ubiquitination. Although both Gag and ESCRTs undergo some level of ubiquitination, definitive demonstration that ubiquitin is required for viral release is lacking. Here we suppress ubiquitination at viral budding sites by fusing the catalytic domain of the Herpes Simplex UL36 deubiquitinating enzyme (DUb) onto TSG101, Alix, or Gag. Results Expressing DUb-TSG101 suppressed Alix-independent HIV-1 release and viral particles remained tethered to the cell surface. DUb-TSG101 had no effect on budding of MoMLV or EIAV, two retroviruses that rely on the ESCRT machinery for exit. Alix-dependent virus release such as EIAV’s, and HIV-1 lacking access to TSG101, was instead dramatically blocked by co-expressing DUb-Alix. Finally, Gag-DUb was unable to support virus release and dominantly interfered with release of wild type HIV-1. Fusion of UL36 did not effect interactions with Alix, TSG101, or Gag and all of the inhibitory effects of UL36 fusion were abolished when its catalytic activity was ablated. Accordingly, Alix, TSG101 and Gag fused to inactive UL36 functionally replaced their unfused counterparts. Interestingly, coexpression of the Nedd4-2s ubiquitin ligase suppressed the ability of DUb-TSG101 to inhibit HIV-1 release while also restoring detectable Gag ubiquitination at the membrane. Similarly, incorporation of Gag-Ub fusion proteins into virions lifted DUb-ESCRT inhibitory effect. In contrast, Nedd4-2s did not suppress the inhibition mediated by Gag-DUb despite restoring robust ubiquitination of TSG101/ESCRT-I at virus budding sites. Conclusions These studies demonstrate a necessary and

  12. Predictive Significance of Tumor Depth and Budding for Late Lymph Node Metastases in Patients with Clinical N0 Early Oral Tongue Carcinoma.

    Science.gov (United States)

    Hori, Yukiko; Kubota, Akira; Yokose, Tomoyuki; Furukawa, Madoka; Matsushita, Takeshi; Takita, Morihito; Mitsunaga, Sachiyo; Mizoguchi, Nobutaka; Nonaka, Tetsuo; Nakayama, Yuko; Oridate, Nobuhiko

    2017-04-03

    In clinical N0 early oral tongue carcinoma, treatment of occult lymph node metastasis is controversial. The purpose of this study was to assess the histopathological risk factors for predicting late lymph node metastasis in early oral tongue carcinoma. We retrospectively reviewed 48 patients with early oral tongue squamous cell carcinoma. Associations between the histopathological factors (depth of tumor, differentiation, blood vessel invasion, lymphatic invasion, and tumor budding) and late lymph metastasis were analyzed. Although the univariate analysis identified blood vessel invasion, lymphatic invasion, and high-grade tumor budding as predictive factors for neck recurrence (p tumor budding as an independent predictive factor (p tumor depth ≥ 3 mm and high-grade tumor budding yielded high diagnostic accuracy. Tumor depth and budding grade were identified as histopathological risk factors for late neck recurrence in clinical N0 early oral tongue carcinoma.

  13. Effect of gamma irradiated parenchyma on the growth of irradiated potato tuber buds; Efecto del parenquina irradiado sobre el desarrollo de las yemas de tuberculos de patata tratados por radiacion GAMMA

    Energy Technology Data Exchange (ETDEWEB)

    Fernandez Gonzalez, J.; Garcia Collantes, M. A.

    1976-07-01

    The development of buds greffed on irradiated potato parenchyma was studied. The irradiated parenchyma does not influence the sprouting capacity of buds, but it affects the way they develop. (Author) 9 refs.

  14. Effects of bud load on quality of Beogradska besemena and Thompson seedless table grapes and cultivar differentiation based on chemometrics of analytical indices.

    Science.gov (United States)

    Baiano, Antonietta; Terracone, Carmela

    2012-02-01

    The effects of bud load on the quality of Beogradska besemena and Thompson seedless table grape cultivars were studied. Two pruning treatments were imposed: 44 and 18 buds per vine for Beogradska besemena and 44 and 22 buds per vine for Thompson seedless. In Beogradska besemena the reduction of bud load decreased titratable acidity (-4%), skin dry weight (-16%) and malic and citric acid contents (-43 and - 20%) and increased tartaric acid content (14%). The decrease in pulp antioxidant activity (-36%) was related to the decrease in hydroxycinnamoyl tartaric acid content (-13%). Concerning skin, the reduction of bud load decreased catechin and caffeoyl tartaric acid contents (-42 and - 40%) and significantly increased rutin and quercetin-3-O-glucopyranoside contents as specific compounds and flavonoids and proanthocyanidins (9 and 21%) as classes of compounds, thus causing an increase in antioxidant activity (6%). In Thompson seedless the reduction of bud load increased soluble solid content (7%), acidity (9%) and concentrations of the three organic acids (7, 3 and 14%). The increase in pulp antioxidant activity (25%) could be attributed to the increase in total phenolics (69%). The reduction of bud load caused a significant decrease in quercetin-3-O-glucopyranoside (-26%) and antioxidant activity (-15%) in skin. Principal component analysis allowed good separation between samples of the two cultivars, independently of bud level. The results indicate that the effects of different bud loads are cultivar-dependent. Bud load and genotype differences were shown to dramatically impact the quality and antioxidant properties of table grape. Copyright © 2011 Society of Chemical Industry.

  15. Dormancy of 'Imperial Gala' apple and 'Hosui' pear tree buds in a region of low chill occurrence

    Directory of Open Access Journals (Sweden)

    Ruy Inácio Neiva de Carvalho

    2014-08-01

    Full Text Available The objective of this work was to evaluate the dormancy dynamic of Imperial Gala apple tree buds and Hosui pear tree buds in a region of low chill occurrence. Experiments were conducted between April and August in 2007 and 2008. Branches were collected every two weeks from an orchard at Porto Amazonas (Paraná State, Brazil. On the last sampling day, an additional set of branches was collected and refrigerated between 4°C and 7°C for 1,440 hours. Dormancy was evaluated using a biological test of single node cuttings isolated in growth chambers (GC at 25°C with 16 hours of light exposure. The number of chill hours (CH and chill units (CU for the region were recorded. The two species were evaluated in separate experiments. We used 11 completely randomized treatments with four replicas for each species. The peak of endodormancy for the Imperial Gala apple tree buds occurred in early June 2007 and from middle June to early July in 2008. The endodormancy of the Hosui pear tree buds oscillated between April and August in 2007 and peaked between June and early July in 2008.

  16. Root temperature effects on growth and bud break of Rosa hybrida in relation to cytokinin concentrations in xylem sap.

    NARCIS (Netherlands)

    Dieleman, J.A.; Verstappen, F.W.A.; Kuiper, D.

    1998-01-01

    The effects of three divergent root temperatures (11°C, 20°C and 26°C) on growth and bud break of Rosa hybrida were studied. Root morphology was changed considerably with root temperature. Roots at 11°C were white, succulent, short and sparsely branched, whereas at 26°C roots were long, brown, thin

  17. Tbx2 terminates shh/fgf signaling in the developing mouse limb bud by direct repression of gremlin1

    NARCIS (Netherlands)

    Farin, Henner F.; Lüdtke, Timo H.-W.; Schmidt, Martina K.; Placzko, Susann; Schuster-Gossler, Karin; Petry, Marianne; Christoffels, Vincent M.; Kispert, Andreas

    2013-01-01

    Vertebrate limb outgrowth is driven by a positive feedback loop that involves Sonic hedgehog (Shh) and Gremlin1 (Grem1) in the posterior limb bud mesenchyme and Fibroblast growth factors (Fgfs) in the overlying epithelium. Proper spatio-temporal control of these signaling activities is required to

  18. Viruses budding from either the apical or the basolateral plasma membrane domain of MDCK cells have unique phospholipid compositions

    NARCIS (Netherlands)

    van Meer, G.; Simons, K.

    1982-01-01

    Influenza virus and vesicular stomatitis virus (VSV) obtain their lipid envelope by budding through the plasma membrane of infected cells. When monolayers of Madin-Darby canine kidney (MDCK) cells, a polarized epithelial cell line, are infected with fowl plague virus (FPV), an avian influenza virus,

  19. Update History of This Database - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available switchLanguage; BLAST Search Image Search Home About Archive Update History Data ...List Contact us Budding yeast cDNA sequencing project Update History of This Database Date Update contents 2...cription Download License Update History of This Database Site Policy | Contact Us Update Histor

  20. Migration Monitoring of Blackcurrant Gall Mite (Cecidophyopsis ribis Westw. from Buds to Leaves on Several Blackcurrant (Ribes nigrum L. Cultivars

    Directory of Open Access Journals (Sweden)

    Piotrowski Wojciech

    2016-12-01

    Full Text Available The blackcurrant gall mite (Cecidophyopsis ribis is the most important pest of blackcurrant crops. Over recent years withdrawal from plant protection programmes of chemical products (endosulfan and amitraz used for the control of this pest in Poland, has led to an observed increase in population numbers. In 2013, fenpiroxymate (Ortus 05 SC became registered for control of this pest. It is deemed best that chemical protection should be used during the migration period; when big gall mites emerge from buds in search of new buds. The studies were carried out in a plantation of blackcurrants during 2011-2013. The assessment of migration of the blackcurrant gall mite was carried out on the cultivars ‘Ben Hope’, ‘Ben Alde’r, ‘Ojeby’n and ‘Ruben’. Every year, from selected cultivars buds were collected. They were then placed on blackcurrant leaves within Petri dishes. After one, three and five days of placing buds on the leaves, the estimated number of eriophyid mites on the leaves was calculated. The data has shown a very useful method for monitoring blackcurrant gall mite, which can be used in calculating the treatment dates for this pest. Also, the data has shown that differences in the periods of migration of the mite are dependent on the cultivar and time of flowering. Among the cultivars observed the least susceptible to colonization by the blackcurrant gall mite was a Polish cultivar ‘Ruben’, while the most susceptible cultivar was ‘Ben Hope’.

  1. Tumor Budding, Micropapillary Pattern, and Polyploidy Giant Cancer Cells in Colorectal Cancer: Current Status and Future Prospects

    Directory of Open Access Journals (Sweden)

    Shiwu Zhang

    2016-01-01

    Full Text Available We previously reported that polyploid giant cancer cells (PGCGs induced by CoCl2 could form through endoreduplication or cell fusion. A single PGCC formed tumors in immunodeficient mice. PGCCs are also the key contributors to the cellular atypia and associate with the malignant grade of tumors. PGCCs have the properties of cancer stem cells and produce daughter cells via asymmetric cell division. Compared with diploid cancer cells, these daughter cells express less epithelial markers and acquire mesenchymal phenotype with importance in cancer development and progression. Tumor budding is generally recognized to correlate with a high recurrence rate, lymph node metastasis, chemoresistance, and poor prognosis of colorectal cancers (CRCs and is a good indicator to predict the metastasis and aggressiveness in CRCs. Micropapillary pattern is a special morphologic pattern and also associates with tumor metastasis and poor prognosis. There are similar morphologic features and molecular phenotypes among tumor budding, micropapillary carcinoma pattern, and PGCCs with their budding daughter cells and all of them show strong ability of tumor invasion and migration. In this review, we discuss the cancer stem cell properties of PGCCs, the molecular mechanisms of their regulation, and the relationships with tumor budding and micropapillary pattern in CRCs.

  2. Tumor Budding, Micropapillary Pattern, and Polyploidy Giant Cancer Cells in Colorectal Cancer: Current Status and Future Prospects

    Science.gov (United States)

    Zhang, Dan; Yang, Zhengduo; Zhang, Xipeng

    2016-01-01

    We previously reported that polyploid giant cancer cells (PGCGs) induced by CoCl2 could form through endoreduplication or cell fusion. A single PGCC formed tumors in immunodeficient mice. PGCCs are also the key contributors to the cellular atypia and associate with the malignant grade of tumors. PGCCs have the properties of cancer stem cells and produce daughter cells via asymmetric cell division. Compared with diploid cancer cells, these daughter cells express less epithelial markers and acquire mesenchymal phenotype with importance in cancer development and progression. Tumor budding is generally recognized to correlate with a high recurrence rate, lymph node metastasis, chemoresistance, and poor prognosis of colorectal cancers (CRCs) and is a good indicator to predict the metastasis and aggressiveness in CRCs. Micropapillary pattern is a special morphologic pattern and also associates with tumor metastasis and poor prognosis. There are similar morphologic features and molecular phenotypes among tumor budding, micropapillary carcinoma pattern, and PGCCs with their budding daughter cells and all of them show strong ability of tumor invasion and migration. In this review, we discuss the cancer stem cell properties of PGCCs, the molecular mechanisms of their regulation, and the relationships with tumor budding and micropapillary pattern in CRCs. PMID:27843459

  3. Tumor cell budding from focally disrupted tumor capsules: a common pathway for all breast cancer subtype derived invasion?

    Directory of Open Access Journals (Sweden)

    Yan-gao Man

    2010-01-01

    Full Text Available Human breast cancer represents a group of highly heterogeneous lesions consisting of about 20 morphologically and immnohistochemically distinct subtypes with substantially different prognoses. Our recent studies have suggested that all breast cancer subtypes, however, may share a common pathway, tumor cell budding from focally disrupted tumor capsules, for their invasion. The potential mechanisms and clinical implications of our observations are discussed.

  4. The tumor microenvironment: An irreplaceable element of tumor budding and epithelial-mesenchymal transition-mediated cancer metastasis.

    Science.gov (United States)

    Li, Hui; Xu, Fangying; Li, Si; Zhong, Anjing; Meng, Xianwen; Lai, Maode

    2016-07-03

    Tumor budding occurs at the invasive front of cancer; the tumor cells involved have metastatic and stemness features, indicating a poor prognosis. Tumor budding is partly responsible for cancer metastasis, and its initiation is based on the epithelial-mesenchymal transition (EMT) process. The EMT process involves the conversion of epithelial cells into migratory and invasive cells, and is a profound event in tumorigenesis. The EMT, associated with the formation of cancer stem cells (CSCs) and resistance to therapy, results from a combination of gene mutation, epigenetic regulation, and microenvironmental control. Tumor budding can be taken to represent the EMT in vivo. The EMT process is under the influence of the tumor microenvironment as well as tumor cells themselves. Here, we demonstrate that the tumor microenvironment dominates EMT development and impacts cancer metastasis, as well as promotes CSC formation and mediates drug resistance. In this review, we mainly discuss components of the microenvironment, such as the extracellular matrix (ECM), inflammatory cytokines, metabolic products, and hypoxia, that are involved in and impact on the acquisition of tumor-cell motility and dissemination, the EMT, metastatic tumor-cell formation, tumor budding and CSCs, and cancer metastasis, including subsequent chemo-resistance. From our point of view, the tumor microenvironment now constitutes a promising target for cancer therapy.

  5. Measurement of the volume growth rate of single budding yeast with the MOSFET-based microfluidic Coulter counter.

    Science.gov (United States)

    Sun, Jiashu; Stowers, Chris C; Boczko, Erik M; Li, Deyu

    2010-11-07

    We report on measurements of the volume growth rate of ten individual budding yeast cells using a recently developed MOSFET-based microfluidic Coulter counter. The MOSFET-based microfluidic Coulter counter is very sensitive, provides signals that are immune from the baseline drift, and can work with cell culture media of complex composition. These desirable features allow us to directly measure the volume growth rate of single cells of Saccharomyces cerevisiae LYH3865 strain budding yeast in YNB culture media over a whole cell cycle. Results indicate that all budding yeast follow a sigmoid volume growth profile with reduced growth rates at the initial stage before the bud emerges and the final stage after the daughter gets mature. Analysis of the data indicates that even though all piecewise linear, Gomperitz, and Hill's function models can fit the global growth profile equally well, the data strongly support local exponential growth phenomenon. Accurate volume growth measurements are important for applications in systems biology where quantitative parameters are required for modeling and simulation.

  6. A method for labeling proteins with tags at the native genomic loci in budding yeast.

    Directory of Open Access Journals (Sweden)

    Qian Wang

    Full Text Available Fluorescent proteins and epitope tags are often used as protein fusion tags to study target proteins. One prevailing technique in the budding yeast Saccharomyces cerevisiae is to fuse these tags to a target gene at the precise chromosomal location via homologous recombination. However, several limitations hamper the application of this technique, such as the selectable markers not being reusable, tagging of only the C-terminal being possible, and a "scar" sequence being left in the genome. Here, we describe a strategy to solve these problems by tagging target genes based on a pop-in/pop-out and counter-selection system. Three fluorescent protein tag (mCherry, sfGFP, and mKikGR and two epitope tag (HA and 3×FLAG constructs were developed and utilized to tag HHT1, UBC13 or RAD5 at the chromosomal locus as proof-of-concept.

  7. Incidence, progression and intensity of Bud Rot in Elaeis guineensis Jacq. in San Lorenzo, Ecuador

    Directory of Open Access Journals (Sweden)

    Fernando Rivas Figueroa

    2017-01-01

    Full Text Available BUD rot (BR is the most serious disease of oil palm in Latin America; in Equator has caused more than 150 million USD of losses. The aim of this work was to determine the incidence, progression and disease intensity of BR in E. guineensis. Incidence and disease progression was determined from data of oil palm enterprises: Palesema, PDA, Palpailón, Energy & Palma y Alespalma during 2006-2013. Disease intensity was determined at 2013. Incidence was 66.75 % and disease intensity was 46 %. Based on projections of accumulative incidence a polynomial equation was built that predicted 78.30 % of cumulative incidence for 2014, indicating exponential growth of BR from 2009 to 2013. Magnitude of damages based on incidence, disease progression and infection index indicated the occurrence of a lethal form of BR in San Lorenzo, province of Esmeraldas, Equator.

  8. Phosphorylation-Dependent Activation of the ESCRT Function of ALIX in Cytokinetic Abscission and Retroviral Budding.

    Science.gov (United States)

    Sun, Sheng; Sun, Le; Zhou, Xi; Wu, Chuanfen; Wang, Ruoning; Lin, Sue-Hwa; Kuang, Jian

    2016-02-08

    The modular adaptor protein ALIX is a key player in multiple ESCRT-III-mediated membrane remodeling processes. ALIX is normally present in a closed conformation due to an intramolecular interaction that renders ALIX unable to perform its ESCRT functions. Here we demonstrate that M phase-specific phosphorylation of the intramolecular interaction site within the proline-rich domain (PRD) of ALIX transforms cytosolic ALIX from closed to open conformation. Defining the role of this mechanism of ALIX regulation in three classical ESCRT-mediated processes revealed that phosphorylation of the intramolecular interaction site in the PRD is required for ALIX to function in cytokinetic abscission and retroviral budding, but not in multivesicular body sorting of activated epidermal growth factor receptor. Thus, phosphorylation of the intramolecular interaction site in the PRD is one of the major mechanisms that activates the ESCRT function of ALIX. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Bionomy of spruce bud scale, Physokermes piceae (schrank (Hemiptera: Coccidae in the Belgrade area, Serbia

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    Graora Draga

    2012-01-01

    Full Text Available Spruce bud scale, Physokermes piceae, develops on the genus Picea. Large colonies of this species are constantly present on Picea abies in green areas in the Belgrade territory, causing the drying of needles, branches and whole plants. Therefore, Ph. piceae is a significant spruce pest. Spruce scales attract many entomophagous insects able to reduce pest population. Parasitoid wasps Coccophagus lycimnia (Walk (Aphelinidae and Microterys lunatus (Dalm. (Encyrtidae were reared. Predators Exochomus quadripustulatus L., Scymnus abietis Paykull (Coccinellidae and Anthribus nebulosus Forster (Anthribidae were determined. Both species of ladybird were confirmed as predators of Ph. piceae for the first time in Serbia, while S. abietis is a new species for the fauna of Serbia. The most effective natural enemy of Ph. piceae was A. nebulosus, reducing populations by 68-80%.

  10. Bifurcation analysis of a model of the budding yeast cell cycle

    Science.gov (United States)

    Battogtokh, Dorjsuren; Tyson, John J.

    2004-09-01

    We study the bifurcations of a set of nine nonlinear ordinary differential equations that describe regulation of the cyclin-dependent kinase that triggers DNA synthesis and mitosis in the budding yeast, Saccharomyces cerevisiae. We show that Clb2-dependent kinase exhibits bistability (stable steady states of high or low kinase activity). The transition from low to high Clb2-dependent kinase activity is driven by transient activation of Cln2-dependent kinase, and the reverse transition is driven by transient activation of the Clb2 degradation machinery. We show that a four-variable model retains the main features of the nine-variable model. In a three-variable model exhibiting birhythmicity (two stable oscillatory states), we explore possible effects of extrinsic fluctuations on cell cycle progression.

  11. Identification of cellular factors required for the budding of koala retrovirus.

    Science.gov (United States)

    Shimode, Sayumi; Nakaoka, Rie; Hoshino, Shigeki; Abe, Masumi; Shogen, Hiroko; Yasuda, Jiro; Miyazawa, Takayuki

    2013-07-01

    Koala retrovirus (KoRV) is a unique gammaretrovirus that is currently endogenizing into its host and considered to be associated with leukemia, lymphoma and immunosuppression in koalas (Phascolactos cinereus). In this study, it was demonstrated that WWP2 or WWP2-like E3 ubiquitin ligases possessing the WW domain closely related to WWP2 and Vps4A/B are involved in KoRV budding. These data suggest that KoRV Gag recruits the cellular endosomal sorting complex required for transport machinery through interaction of the PPPY L-domain with the WW domain(s) of WWP2 and that progeny virions are released from cells by utilizing the multivesicular body sorting pathway. © 2013 The Societies and Wiley Publishing Asia Pty Ltd.

  12. Chromatin-dependent and -independent regulation of DNA replication origin activation in budding yeast.

    Science.gov (United States)

    Lõoke, Marko; Kristjuhan, Kersti; Värv, Signe; Kristjuhan, Arnold

    2013-02-01

    To elucidate the role of the chromatin environment in the regulation of replication origin activation, autonomously replicating sequences were inserted into identical locations in the budding yeast genome and their activation times in S phase determined. Chromatin-dependent origins adopt to the firing time of the surrounding locus. In contrast, the origins containing two binding sites for Forkhead transcription factors are activated early in the S phase regardless of their location in the genome. Our results also show that genuinely late-replicating parts of the genome can be converted into early-replicating loci by insertion of a chromatin-independent early replication origin, ARS607, whereas insertion of two Forkhead-binding sites is not sufficient for conversion.

  13. Glucosylated caffeoylquinic acid derivatives from the flower buds of Lonicera japonica

    Directory of Open Access Journals (Sweden)

    Yang Yu

    2015-05-01

    Full Text Available Three new glucosylated caffeoylquinic acid isomers (1–3, along with six known compounds, have been isolated from an aqueous extract of the flower buds of Lonicera japonica. Structures of the new compounds were determined by spectroscopic and chemical methods as (−-4-O-(4-O-β-d-glucopyranosylcaffeoylquinic acid (1, (−-3-O-(4-O-β-d-glucopyranosylcaffeoylquinic acid (2, and (−-5-O-(4-O-β-d-glucopyranosylcaffeoylquinic acid (3, respectively. In the preliminary in vitro assays, two known compounds methyl caffeate and 2ʹ-O-methyladenosine showed inhibitory activity against Coxsackie virus B3 with IC50 values of 3.70 μmol/L and 6.41 μmol/L and SI values of 7.8 and 12.1, respectively.

  14. Filovirus proteins for antiviral drug discovery: Structure/function of proteins involved in assembly and budding.

    Science.gov (United States)

    Martin, Baptiste; Reynard, Olivier; Volchkov, Viktor; Decroly, Etienne

    2018-02-01

    There are no approved medications for the treatment of Marburg or Ebola virus infection. In two previous articles (Martin et al., 2016, Martin et al., 2017), we reviewed surface glycoprotein and replication proteins structure/function relationship to decipher the molecular mechanisms of filovirus life cycle and identify antiviral strategies. In the present article, we recapitulate knowledge about the viral proteins involved in filovirus assembly and budding. First we describe the structural data available for viral proteins associated with virus assembly and virion egress and then, we integrate the structural features of these proteins in the functional context of the viral replication cycle. Finally, we summarize recent advances in the development of innovative antiviral strategies to target filovirus assembly and egress. The development of such prophylactic or post-exposure treatments could help controlling future filovirus outbreaks. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. A method for labeling proteins with tags at the native genomic loci in budding yeast.

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    Wang, Qian; Xue, Huijun; Li, Siqi; Chen, Ying; Tian, Xuelei; Xu, Xin; Xiao, Wei; Fu, Yu Vincent

    2017-01-01

    Fluorescent proteins and epitope tags are often used as protein fusion tags to study target proteins. One prevailing technique in the budding yeast Saccharomyces cerevisiae is to fuse these tags to a target gene at the precise chromosomal location via homologous recombination. However, several limitations hamper the application of this technique, such as the selectable markers not being reusable, tagging of only the C-terminal being possible, and a "scar" sequence being left in the genome. Here, we describe a strategy to solve these problems by tagging target genes based on a pop-in/pop-out and counter-selection system. Three fluorescent protein tag (mCherry, sfGFP, and mKikGR) and two epitope tag (HA and 3×FLAG) constructs were developed and utilized to tag HHT1, UBC13 or RAD5 at the chromosomal locus as proof-of-concept.

  16. Small bud of probable gastrointestinal stromal tumor within a laparoscopically-resected gastric schwannoma.

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    Cho, Haruhiko; Watanabe, Takafumi; Aoyama, Toru; Hayashi, Tsutomu; Yamada, Takanobu; Ogata, Takashi; Yoshikawa, Takaki; Tsuburaya, Akira; Sekiguchi, Hironobu; Nakamura, Yoshiyasu; Sakuma, Yuji; Kameda, Yoichi; Miyagi, Yohei

    2012-06-01

    Submucosal tumors (SMTs) of the gastrointestinal (GI) tract can be potentially difficulty to diagnose pathologically. We report a case of a gastric SMT that was resected by laparoscopic partial gastrectomy. Although the initial histological and immunohistochemical examinations considered the tumor as a schwannoma, mRNA-based KIT genotyping indicated that the tumor included cells with KIT gene expression, and that a small number of cells carried a deletion mutation in exon 11. Additional histopathological investigations revealed small aggregates of enlarged spindle to epithelioid cells, which were positive for KIT, CD34 and DOG1, and negative for S-100, scattered among the S-100-positive schwannoma cells. We consider that the cells carrying the KIT gene mutation are microscopic buds of a gastrointestinal stroma tumor (GIST), and to the best of our knowledge, this is the first report of probable GIST tissues identified in a schwannoma. Our observations raised the significance of genotyping for diagnosis of GI tract SMTs.

  17. Evidence for a role of glutamate as an efferent transmitter in taste buds

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    Anderson Catherine B

    2010-06-01

    Full Text Available Abstract Background Glutamate has been proposed as a transmitter in the peripheral taste system in addition to its well-documented role as an umami taste stimulus. Evidence for a role as a transmitter includes the presence of ionotropic glutamate receptors in nerve fibers and taste cells, as well as the expression of the glutamate transporter GLAST in Type I taste cells. However, the source and targets of glutamate in lingual tissue are unclear. In the present study, we used molecular, physiological and immunohistochemical methods to investigate the origin of glutamate as well as the targeted receptors in taste buds. Results Using molecular and immunohistochemical techniques, we show that the vesicular transporters for glutamate, VGLUT 1 and 2, but not VGLUT3, are expressed in the nerve fibers surrounding taste buds but likely not in taste cells themselves. Further, we show that P2X2, a specific marker for gustatory but not trigeminal fibers, co-localizes with VGLUT2, suggesting the VGLUT-expressing nerve fibers are of gustatory origin. Calcium imaging indicates that GAD67-GFP Type III taste cells, but not T1R3-GFP Type II cells, respond to glutamate at concentrations expected for a glutamate transmitter, and further, that these responses are partially blocked by NBQX, a specific AMPA/Kainate receptor antagonist. RT-PCR and immunohistochemistry confirm the presence of the Kainate receptor GluR7 in Type III taste cells, suggesting it may be a target of glutamate released from gustatory nerve fibers. Conclusions Taken together, the results suggest that glutamate may be released from gustatory nerve fibers using a vesicular mechanism to modulate Type III taste cells via GluR7.

  18. Dilution of the cell cycle inhibitor Whi5 controls budding yeast cell size

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    Schmoller, Kurt M.; Turner, J.J.; Kõivomägi, M.; Skotheim, Jan M.

    2015-01-01

    Cell size fundamentally affects all biosynthetic processes by determining the scale of organelles and influencing surface transport1,2. Although extensive studies have identified many mutations affecting cell size, the molecular mechanisms underlying size control have remained elusive3. In budding yeast, size control occurs in G1 phase prior to Start, the point of irreversible commitment to cell division4,5. It was previously thought that activity of the G1 cyclin Cln3 increased with cell size to trigger Start by initiating the inhibition of the transcriptional inhibitor Whi56-8. However, while Cln3 concentration does modulate the rate at which cells pass Start, we found that its synthesis increases in proportion to cell size so that its total concentration is nearly constant during pre-Start G1. Rather than increasing Cln3 activity, we identify decreasing Whi5 activity — due to the dilution of Whi5 by cell growth — as a molecular mechanism through which cell size controls proliferation. Whi5 is synthesized in S/G2/M phases of the cell cycle in a largely size-independent manner. This results in smaller daughter cells being born with higher Whi5 concentrations that extend their pre-Start G1 phase. Thus, at its most fundamental level, budding yeast size control results from the differential scaling of Cln3 and Whi5 synthesis rates with cell size. More generally, our work shows that differential size-dependency of protein synthesis can provide an elegant mechanism to coordinate cellular functions with growth. PMID:26390151

  19. Daughter-specific transcription factors regulate cell size control in budding yeast.

    Directory of Open Access Journals (Sweden)

    Stefano Di Talia

    2009-10-01

    Full Text Available In budding yeast, asymmetric cell division yields a larger mother and a smaller daughter cell, which transcribe different genes due to the daughter-specific transcription factors Ace2 and Ash1. Cell size control at the Start checkpoint has long been considered to be a main regulator of the length of the G1 phase of the cell cycle, resulting in longer G1 in the smaller daughter cells. Our recent data confirmed this concept using quantitative time-lapse microscopy. However, it has been proposed that daughter-specific, Ace2-dependent repression of expression of the G1 cyclin CLN3 had a dominant role in delaying daughters in G1. We wanted to reconcile these two divergent perspectives on the origin of long daughter G1 times. We quantified size control using single-cell time-lapse imaging of fluorescently labeled budding yeast, in the presence or absence of the daughter-specific transcriptional regulators Ace2 and Ash1. Ace2 and Ash1 are not required for efficient size control, but they shift the domain of efficient size control to larger cell size, thus increasing cell size requirement for Start in daughters. Microarray and chromatin immunoprecipitation experiments show that Ace2 and Ash1 are direct transcriptional regulators of the G1 cyclin gene CLN3. Quantification of cell size control in cells expressing titrated levels of Cln3 from ectopic promoters, and from cells with mutated Ace2 and Ash1 sites in the CLN3 promoter, showed that regulation of CLN3 expression by Ace2 and Ash1 can account for the differential regulation of Start in response to cell size in mothers and daughters. We show how daughter-specific transcriptional programs can interact with intrinsic cell size control to differentially regulate Start in mother and daughter cells. This work demonstrates mechanistically how asymmetric localization of cell fate determinants results in cell-type-specific regulation of the cell cycle.

  20. Dilution of the cell cycle inhibitor Whi5 controls budding-yeast cell size.

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    Schmoller, Kurt M; Turner, J J; Kõivomägi, M; Skotheim, Jan M

    2015-10-08

    Cell size fundamentally affects all biosynthetic processes by determining the scale of organelles and influencing surface transport. Although extensive studies have identified many mutations affecting cell size, the molecular mechanisms underlying size control have remained elusive. In the budding yeast Saccharomyces cerevisiae, size control occurs in G1 phase before Start, the point of irreversible commitment to cell division. It was previously thought that activity of the G1 cyclin Cln3 increased with cell size to trigger Start by initiating the inhibition of the transcriptional inhibitor Whi5 (refs 6-8). Here we show that although Cln3 concentration does modulate the rate at which cells pass Start, its synthesis increases in proportion to cell size so that its total concentration is nearly constant during pre-Start G1. Rather than increasing Cln3 activity, we identify decreasing Whi5 activity--due to the dilution of Whi5 by cell growth--as a molecular mechanism through which cell size controls proliferation. Whi5 is synthesized in S/G2/M phases of the cell cycle in a largely size-independent manner. This results in smaller daughter cells being born with higher Whi5 concentrations that extend their pre-Start G1 phase. Thus, at its most fundamental level, size control in budding yeast results from the differential scaling of Cln3 and Whi5 synthesis rates with cell size. More generally, our work shows that differential size-dependency of protein synthesis can provide an elegant mechanism to coordinate cellular functions with growth.